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- | <b>Rebekka Bauer</b><br> | + | |
- | Action points:<br>
| + | <b>Action points for the day:</b><br> |
| -bios on the wiki<br> | | -bios on the wiki<br> |
| -sci-fi meeting<br> | | -sci-fi meeting<br> |
| -characterisation talk<br> | | -characterisation talk<br> |
| -come up with more ideas<br> | | -come up with more ideas<br> |
- | -do more research on anti-venom and prodigiosin<br> | + | -do more research on anti-venom and prodigiosin<br><br> |
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- | <br><br>
| + | We started the day off by brainstorming several idea and selecting the ones we thought were viable enough to be presented to the professors on Friday.<br><br> |
- | Sci-Fi prodigiosin ideas (developed with Nick Kral and CJ from the RCA):<br>
| + | <b>Brainstorming:</b><br> |
- | -use prodigiosin (red pigment) as the new "colour of health" (know something is sterile rather than assume it is)<br>
| + | |
- | -possible future uses: in decontamination/ as a "panic room"/ sterile hospice<br>
| + | |
- | -possible future products: hand gel, clothing (e.g. protective suits in bioreactor plants), decontamination paint (in hospitals etc)<br>
| + | |
- | -actual uses: anti-cancer (could be in a red drip, red pill?), anti-malarial (drug verification because pigment colour is hard to fake)<br>
| + | |
- | Eventually, this idea was scratched because optimising the production pathway does not contain enough synthetic biology. In addition, the compound is immunosuppressive (http://pubs.rsc.org/en/Content/ArticleHtml/2008/CC/b719353j) and would therefore be disadvantageous to normal people. Many of the envisaged applications would only work with less problematic analogues of prodigiosin.
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- | <br><br> | + | |
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- | Brainstorming:<br> | + | |
| Venom and BAS ideas to be presented on Friday!<br> | | Venom and BAS ideas to be presented on Friday!<br> |
| -desertification (overdone already)<br> | | -desertification (overdone already)<br> |
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| -serum bile acid as a biomarker of liver problems in pregnancy<br><br> | | -serum bile acid as a biomarker of liver problems in pregnancy<br><br> |
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- | Research for venom:<br>
| + | After the brain-storming session we had a talk from Chris about characterisation giving me a glimpse into what I would be expecting in the weeks after the project has been pinned down on a single idea. <br><br> |
- | organise slides into problem, specifications needed to tackle problem, how to achieve specifications<br>
| + | <b>Characterization presentation by Chris:</b><br> |
- | -venom chosen: Asian cobra (one of the most common ones)<br>
| + | in vivo presentation<br> |
- | -Ming: summarise problem and specifications needed<br>
| + | -Timer vs. switch. need to know characteristics of parts. <br> |
- | -Chris: look at shark antibodies<br>
| + | -How to characterise: Look at individual parts. What goes in, what goes out. What function is carried out. <br> |
- | -Nikki: look at in vivo mutagenesis (PCR mutagenesis would be too tedious)<br>
| + | -What specifically do you want to characterise? Terminator, core-promoter, RBS, reporter, coding region. <br> |
- | -Frank: selection (FACS with GFP?)<br>
| + | -Terminator requires several different characterizations. <br> |
- | -Rebekka: genetic circuit (detection mechanism - two component system?, suicide mechanism for cells that do not detect venom)
| + | -Place promoter, coding region into an expression circuit with a reporter. <br> |
- | | + | -Standards are only existant for promoters. How strong promoters are. <br> |
- | | + | -Canton paper 2008: Base characteristics that you need to know about an inducible promoter. <br> |
- | <html>
| + | -How responsive (transfer function of response), dynamic? (output) All relative to J23101. <br> |
- | <p>
| + | -Synthesis rate, number of cells and divide your promoter output with standard which gives you a value relative to J23101. RPU (relative promoter unit) <br> |
- | <p>
| + | -RBS calculator gives you a value to how strong it is. Need to debug yourself. <br> |
- | <DIV style="TEXT-INDENT: -7.1pt; MARGIN: 0cm 0cm 0pt 7.1pt; mso-char-indent-count: 0; mso-list: l3 level1 lfo4" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-bidi-font-size: 11.0pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=5 face=Calibri><STRONG></STRONG></FONT></SPAN></SPAN></FONT> </DIV>
| + | -Hard to characterise until we know what we want to do. <br> |
- | <DIV style="TEXT-INDENT: -7.1pt; MARGIN: 0cm 0cm 0pt 7.1pt; mso-char-indent-count: 0; mso-list: l3 level1 lfo4" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-bidi-font-size: 11.0pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=5 face=Calibri><STRONG>Si, Nina, Nick and Yuanwei</STRONG></FONT></SPAN></SPAN></FONT></DIV>
| + | -Plate readers look at OD, fluorescene, luminescence. <br> |
- | <DIV style="TEXT-INDENT: -7.1pt; MARGIN: 0cm 0cm 0pt 7.1pt; mso-char-indent-count: 0; mso-list: l3 level1 lfo4" class=MsoListParagraph><FONT color=#000000><SPAN style="FONT-SIZE: 14pt; mso-bidi-font-size: 11.0pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT face=Calibri></FONT></SPAN></SPAN></FONT> </DIV>
| + | -Individual cells analyzed for GFP and RFP. <br> |
- | <DIV style="TEXT-INDENT: -7.1pt; MARGIN: 0cm 0cm 0pt 7.1pt; mso-char-indent-count: 0; mso-list: l3 level1 lfo4" class=MsoListParagraph><FONT color=#000000><SPAN style="FONT-SIZE: 14pt; mso-bidi-font-size: 11.0pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><B style="mso-bidi-font-weight: normal"><SPAN style="FONT-SIZE: 14pt; mso-bidi-font-size: 11.0pt" lang=EN-US><FONT face=Calibri>morning:<?xml:namespace prefix = o ns = "urn:schemas-microsoft-com:office:office" /><o:p></o:p></FONT></SPAN></B></FONT></DIV>
| + | Chris works on methods to make it more high-throughput (automates to make faster). Won't consider optimizing something unless it worked twice on the assays. |
- | <P style="TEXT-INDENT: 5.25pt; MARGIN: 0cm 0cm 0pt; mso-char-indent-count: .5" class=MsoNormal><SPAN lang=EN-US><FONT color=#000000 size=3 face=Calibri>brain storming:</FONT></SPAN></P>
| + | Fluorescence doesn't give use pinpoint accuracy. Can't be measured really well, only bulk events and not specific events. |
- | <P style="TEXT-INDENT: 5.25pt; MARGIN: 0cm 0cm 0pt; mso-char-indent-count: .5" class=MsoNormal><SPAN lang=EN-US><o:p><FONT color=#000000 size=3 face=Calibri> </FONT></o:p></SPAN></P>
| + | Enzymatic activity more done in vitro. You're working with factories that work depending on what they're in. <br> |
- | <P style="TEXT-INDENT: -14.2pt; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0; mso-list: l1 level1 lfo3" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><SPAN lang=EN-US><FONT size=3 face=Calibri>meeting with RCA people </FONT></SPAN></FONT></P>
| + | Can find Chris though James at any time. <br> |
- | <P style="TEXT-INDENT: 0cm; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0" class=MsoListParagraph><SPAN lang=EN-US><o:p><FONT color=#000000 size=3 face=Calibri> </FONT></o:p></SPAN></P>
| + | Make sure positive and negative control will work. <br> |
- | <P style="TEXT-INDENT: -14.2pt; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0; mso-list: l1 level1 lfo3" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><SPAN lang=EN-US><FONT size=3 face=Calibri>presenting the DNA hard drive idea</FONT></SPAN></FONT></P>
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- | <P style="TEXT-INDENT: -21.25pt; MARGIN: 0cm 0cm 0pt 35.45pt; mso-char-indent-count: 0; mso-list: l0 level1 lfo1" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-fareast-font-family: Calibri; mso-bidi-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-bidi-theme-font: minor-latin" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>1.</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><SPAN lang=EN-US><FONT size=3 face=Calibri>use DNA sequence as a medium to store information</FONT></SPAN></FONT></P>
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- | <P style="TEXT-INDENT: 36.75pt; MARGIN: 0cm 0cm 0pt; mso-char-indent-count: 3.5" class=MsoNormal><SPAN lang=EN-US><FONT color=#000000 size=3 face=Calibri>DNA base - quaternary bit - original text </FONT></SPAN></P>
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- | <P style="TEXT-INDENT: -21.25pt; MARGIN: 0cm 0cm 0pt 35.45pt; mso-char-indent-count: 0; mso-list: l0 level1 lfo1" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-fareast-font-family: Calibri; mso-bidi-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-bidi-theme-font: minor-latin" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>2.</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><SPAN lang=EN-US><FONT size=3 face=Calibri>put “tag in” information (i.e some specific base pairs as “prototypes”) to classify the information</FONT></SPAN></FONT></P>
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- | <P style="TEXT-INDENT: -21.25pt; MARGIN: 0cm 0cm 0pt 35.45pt; mso-char-indent-count: 0; mso-list: l0 level1 lfo1" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-fareast-font-family: Calibri; mso-bidi-font-family: Calibri; mso-fareast-theme-font: minor-latin; mso-bidi-theme-font: minor-latin" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>3.</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><SPAN lang=EN-US><FONT size=3 face=Calibri>writing functions in programming language with DNA sequences, use DNA as the function variables</FONT></SPAN></FONT></P>
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- | <P style="TEXT-INDENT: 35.9pt; MARGIN: 0cm 0cm 0pt; mso-char-indent-count: 3.42" class=MsoNormal><SPAN lang=EN-US><o:p><FONT color=#000000 size=3 face=Calibri> </FONT></o:p></SPAN></P>
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- | <P style="MARGIN: 0cm 0cm 0pt -0.1pt; mso-para-margin-left: -.01gd" class=MsoNormal><SPAN lang=EN-US><FONT color=#000000 size=3 face=Calibri>- Sci-Fi:</FONT></SPAN></P>
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- | <P style="TEXT-INDENT: -10.5pt; MARGIN: 0cm 0cm 0pt 10.5pt; mso-char-indent-count: -1.0" class=MsoNormal><SPAN lang=EN-US><FONT color=#000000 size=3 face=Calibri> a. in the future, a bacteria charm/necklace can be made to store and carry the information, such as exam syllabus, genetic disease history, family photos, etc</FONT></SPAN></P>
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- | <P style="TEXT-INDENT: -10.5pt; MARGIN: 0cm 0cm 0pt 10.5pt; mso-char-indent-count: -1.0" class=MsoNormal><FONT size=3><FONT face=Calibri><FONT color=#000000><SPAN lang=EN-US> b. different coding methods and types of bacteria can be chosen by the clients for different levels of security needs ,(some high risky information can be store in </SPAN><SPAN class=st1><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US>Bacillus anthraci for military use)</SPAN><SPAN lang=EN-US><o:p></o:p></SPAN></SPAN></FONT></FONT></FONT></P>
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- | <P style="TEXT-INDENT: -10.5pt; MARGIN: 0cm 0cm 0pt 10.5pt; mso-char-indent-count: -1.0" class=MsoNormal><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000> c. data can also be stored in E.coli in the human digestive system, it can be erased by intaking antibiotics<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="TEXT-INDENT: 36.75pt; MARGIN: 0cm 0cm 0pt; mso-char-indent-count: 3.5" class=MsoNormal><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><o:p><FONT color=#000000 size=3 face=Calibri> </FONT></o:p></SPAN></P>
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- | <P style="TEXT-INDENT: -14.2pt; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0; mso-list: l1 level1 lfo3" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-bidi-font-size: 10.5pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><FONT size=3><FONT face=Calibri>practical problem:<o:p></o:p></FONT></FONT></SPAN></FONT></P>
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- | <P style="TEXT-INDENT: 0cm; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0" class=MsoListParagraph><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>we need a bacteria platform to carry out the operation of the data<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="TEXT-INDENT: 0cm; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0" class=MsoListParagraph><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>(I.e: bio-compiler? bio-CPU?) <o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="MARGIN: 0cm 0cm 0pt 15.75pt; mso-para-margin-left: 1.5gd" class=MsoNormal><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>a compiler consists of sets of programs and logic relations, it is theoretically workable, but obviously too difficult to do as a ten-week project<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="TEXT-INDENT: 36.75pt; MARGIN: 0cm 0cm 0pt; mso-char-indent-count: 3.5" class=MsoNormal><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><o:p><FONT color=#000000 size=3 face=Calibri> </FONT></o:p></SPAN></P>
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- | <P style="TEXT-INDENT: -7.1pt; MARGIN: 0cm 0cm 0pt 7.1pt; mso-char-indent-count: 0; mso-list: l1 level1 lfo3" class=MsoListParagraph><FONT color=#000000><SPAN style="FONT-SIZE: 14pt; mso-bidi-font-size: 10.5pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><B style="mso-bidi-font-weight: normal"><SPAN style="FONT-SIZE: 14pt; mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><FONT face=Calibri>afternoon:<o:p></o:p></FONT></SPAN></B></FONT></P>
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- | <P style="TEXT-INDENT: 5.25pt; MARGIN: 0cm 0cm 0pt; mso-char-indent-count: .5" class=MsoNormal><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>brain storming:<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="TEXT-INDENT: 5.25pt; MARGIN: 0cm 0cm 0pt; mso-char-indent-count: .5" class=MsoNormal><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><o:p><FONT color=#000000 size=3 face=Calibri> </FONT></o:p></SPAN></P>
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- | <P style="TEXT-INDENT: -14.2pt; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0; mso-list: l2 level1 lfo2" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-bidi-font-size: 10.5pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><FONT size=3><FONT face=Calibri>since the bio-compiler idea is abandoned, we thought about other ways to store data:<o:p></o:p></FONT></FONT></SPAN></FONT></P>
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- | <P style="TEXT-INDENT: 0cm; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0" class=MsoListParagraph><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><o:p><FONT color=#000000 size=3 face=Calibri> </FONT></o:p></SPAN></P>
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- | <P style="TEXT-INDENT: -14.2pt; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0; mso-list: l2 level1 lfo2" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-bidi-font-size: 10.5pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><FONT size=3><FONT face=Calibri>ROM memory<o:p></o:p></FONT></FONT></SPAN></FONT></P>
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- | <P style="TEXT-INDENT: 15.75pt; MARGIN: 0cm 0cm 0pt; mso-char-indent-count: 1.5" class=MsoNormal><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>8x8 gird to store 64 bits of data<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="MARGIN: 0cm 0cm 0pt 15.75pt; mso-para-margin-left: 1.5gd" class=MsoNormal><SPAN style="mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>the bacteria interaction can be modified to give to outputs, which indicates the two binary states (0 or 1) <o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="TEXT-INDENT: 0cm; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0" class=MsoListParagraph><SPAN style="mso-bidi-font-size: 10.5pt" lang=EN-US><o:p><FONT color=#000000 size=3 face=Calibri> </FONT></o:p></SPAN></P>
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- | <P style="TEXT-INDENT: -14.2pt; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0; mso-list: l2 level1 lfo2" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-bidi-font-size: 10.5pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><SPAN style="mso-bidi-font-size: 10.5pt" lang=EN-US><FONT size=3><FONT face=Calibri>JK flip-flop or synchronized counter<o:p></o:p></FONT></FONT></SPAN></FONT></P>
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- | <P style="TEXT-INDENT: 0cm; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0" class=MsoListParagraph><SPAN style="mso-bidi-font-size: 10.5pt" lang=EN-US><o:p><FONT color=#000000 size=3 face=Calibri> </FONT></o:p></SPAN></P>
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- | <P style="TEXT-INDENT: -14.2pt; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0; mso-list: l2 level1 lfo2" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-bidi-font-size: 10.5pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><SPAN style="mso-bidi-font-size: 10.5pt" lang=EN-US><FONT size=3><FONT face=Calibri>7-segment display<o:p></o:p></FONT></FONT></SPAN></FONT></P>
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- | <P style="MARGIN: 0cm 0cm 0pt 15.75pt; mso-para-margin-left: 1.5gd" class=MsoNormal><SPAN style="mso-bidi-font-size: 10.5pt" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>one bacterium -four input channels-each channel with two states (0 or 1)-nine outputs to give nine numbers (0 to 9)- control the corresponding segments to display the number<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="TEXT-INDENT: -14.2pt; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0; mso-list: l2 level1 lfo2" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-bidi-font-size: 10.5pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><SPAN style="mso-bidi-font-size: 10.5pt" lang=EN-US><FONT size=3><FONT face=Calibri>bacterial minesweeper<o:p></o:p></FONT></FONT></SPAN></FONT></P>
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- | <P style="TEXT-INDENT: 15.75pt; MARGIN: 0cm 0cm 0pt; mso-char-indent-count: 1.5" class=MsoNormal><SPAN style="mso-bidi-font-size: 10.5pt" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>an inhibitor can be considered as a ”mine”<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="MARGIN: 0cm 0cm 0pt 15.75pt; mso-para-margin-left: 1.5gd" class=MsoNormal><SPAN style="mso-bidi-font-size: 10.5pt" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>the main problem is that secondary diffusion is very difficult to control. a mass transfer equation must be modeled for each individual square<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="MARGIN: 0cm 0cm 0pt" class=MsoNormal><SPAN style="mso-bidi-font-size: 10.5pt" lang=EN-US><o:p><FONT color=#000000 size=3 face=Calibri> </FONT></o:p></SPAN></P>
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- | <P style="MARGIN: 0cm 0cm 0pt" class=MsoNormal><B style="mso-bidi-font-weight: normal"><SPAN style="COLOR: #7030a0; FONT-SIZE: 12pt; mso-bidi-font-size: 10.5pt" lang=EN-US><FONT face=Calibri><FONT color=#9900ff>Bile Acid Sensor:<o:p></o:p></FONT></FONT></SPAN></B></P>
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- | <P style="TEXT-INDENT: -14.2pt; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0; mso-list: l2 level1 lfo2" class=MsoListParagraph><FONT color=#000000><SPAN style="mso-bidi-font-size: 10.5pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN><SPAN style="mso-bidi-font-size: 10.5pt" lang=EN-US><FONT size=3><FONT face=Calibri>the idea came up with SI’s final year project<o:p></o:p></FONT></FONT></SPAN></FONT></P>
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- | <P style="TEXT-INDENT: -14.2pt; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0; mso-list: l2 level1 lfo2" class=MsoListParagraph><FONT color=#000000><EM><SPAN style="COLOR: windowtext; mso-bidi-font-size: 10.5pt; mso-ascii-font-family: Calibri; mso-fareast-font-family: Calibri; mso-hansi-font-family: Calibri; mso-bidi-font-family: Calibri" lang=EN-US><SPAN style="mso-list: Ignore"><FONT size=3 face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></SPAN></SPAN></EM><FONT size=3><FONT face=Calibri><SPAN style="mso-bidi-font-size: 10.5pt" lang=EN-US>a biosensor can be produced for daily home use to detect the bile acid </SPAN><SPAN style="COLOR: #0d0d0d; mso-bidi-font-size: 10.5pt; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin; mso-themecolor: text1; mso-themetint: 242" lang=EN-US>concentration level in blood to prevent a series of liver diseases, especially the </SPAN><SPAN><SPAN style="COLOR: #0d0d0d; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin; mso-themecolor: text1; mso-themetint: 242" lang=EN-US>Intrahepatic cholestasis of </SPAN>pregnancy </SPAN><SPAN><SPAN style="COLOR: #0d0d0d; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin; mso-themecolor: text1; mso-themetint: 242" lang=EN-US>(</SPAN>ICP)<o:p></o:p></SPAN></FONT></FONT></FONT></P>
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- | <P style="TEXT-INDENT: -14.2pt; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0; mso-list: l2 level1 lfo2" class=MsoListParagraph><SPAN><FONT color=#000000><SPAN style="mso-list: Ignore"><EM><FONT size=3 face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></EM></SPAN><FONT size=3><FONT face=Calibri>the basic concept behind this sensor:<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="MARGIN: 0cm 0cm 0pt 15.75pt; mso-para-margin-left: 1.5gd" class=MsoNormal><SPAN style="COLOR: #0d0d0d; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin; mso-themecolor: text1; mso-themetint: 242" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>bile acid - enzyme binding with the acid molecules – promoter – triggering the FXR gene – production of GFP<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="TEXT-INDENT: -14.2pt; MARGIN: 0cm 0cm 0pt 14.2pt; mso-char-indent-count: 0; mso-list: l2 level1 lfo2" class=MsoListParagraph><SPAN><FONT color=#000000><SPAN style="mso-list: Ignore"><EM><FONT size=3 face=Calibri>-</FONT><SPAN style="FONT: 7pt 'Times New Roman'"> </SPAN></EM></SPAN><FONT size=3><FONT face=Calibri>GFP is normally used as an indicator of the bio-sensor<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="TEXT-INDENT: 15.75pt; MARGIN: 0cm 0cm 0pt; mso-char-indent-count: 1.5" class=MsoNormal><SPAN style="COLOR: #0d0d0d; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin; mso-themecolor: text1; mso-themetint: 242" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>the main problem is that the fluorescent intensity is very hard to quantified for a home-user<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="TEXT-INDENT: -15.75pt; MARGIN: 0cm 0cm 0pt 15.75pt; mso-char-indent-count: -1.5" class=MsoNormal><SPAN style="COLOR: #0d0d0d; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin; mso-themecolor: text1; mso-themetint: 242" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>- <SPAN style="mso-spacerun: yes"> </SPAN>being inspired by the cosmetic skin colour sample card , we can make a sample card of fluorescence to give the rough concentration level of bile acid<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="TEXT-INDENT: -15.75pt; MARGIN: 0cm 0cm 0pt 15.75pt; mso-char-indent-count: -1.5" class=MsoNormal><SPAN style="COLOR: #0d0d0d; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin; mso-themecolor: text1; mso-themetint: 242" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>- <SPAN style="mso-spacerun: yes"> </SPAN>a threshold value is required to tell the patient when their blood bile acid level is dangerous and may need a medical treatment<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="TEXT-INDENT: -15.75pt; MARGIN: 0cm 0cm 0pt 15.75pt; mso-char-indent-count: -1.5" class=MsoNormal><SPAN style="COLOR: #0d0d0d; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin; mso-themecolor: text1; mso-themetint: 242" lang=EN-US><FONT size=3><FONT face=Calibri><FONT color=#000000>-<SPAN style="mso-spacerun: yes"> </SPAN>therefore, we will modify the linear relationship between bile acid concentration and GFP intensity level into a Hill system using Hill equation to find the threshold value<o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="MARGIN: 0cm 0cm 0pt" class=MsoNormal><SPAN><FONT size=3><FONT face=Calibri><FONT color=#000000>- <SPAN style="mso-spacerun: yes"> </SPAN>also, we may use colour indicator instead of GFP (light indicator) <SPAN style="mso-spacerun: yes"> </SPAN><SPAN style="mso-spacerun: yes"> </SPAN><SPAN style="mso-spacerun: yes"> </SPAN><o:p></o:p></FONT></FONT></FONT></SPAN></P>
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- | <P style="MARGIN: 0cm 0cm 0pt" class=MsoNormal><SPAN style="COLOR: #0d0d0d; mso-bidi-font-family: Calibri; mso-bidi-theme-font: minor-latin; mso-themecolor: text1; mso-themetint: 242" lang=EN-US><o:p><FONT color=#000000 size=3 face=Calibri> </FONT></o:p></SPAN></P>
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- | </html>
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- | <html><br><br><b>Christopher Schoene</b><br><br>
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- | Today we discussed the anti-venom idea with Koby (the RCA student that had arrived). He helped us expand the idea to maybe provide protection against viruses as well. We even developed the idea to use a seasonal hand wash containing the purified antibodies from the season's viruses in order to create a world where a handshake would be more than just a form of greeting but also a way to pass on immunity.<br><br>
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- | | + | |
- | After the brain-storming session we had a talk from Chris about characterisation giving me a glimpse into what I would be expecting in the weeks after the project has been pinned down on a single idea. We currently have two running ideas; the anti-venom and a bile acid sensor. We have split off into two teams and each is working hard in order to build a system that the professors might agree to this Friday at 3:30pm. The deadline is approaching and we still require 2 more possible projects to present to the professors. I will work with my team to complete the anti-venom idea tomorrow and I will try to research a way to improve upon the idea of the use of yeast in water retention by making the express mucins.<br><br> | + | |
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- | RCA sci-fi story ideas: <br>
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- | Nuclear winter leaving people without immune systems. Use external immune system to save everybody? <br><br>
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- | Synthetic life can only use certain amino acids, firewall? <br>
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- | Biocomputers where man and machine are converged closer together. <br>
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- | Floppy disk baby? <br>
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- | Mock news articles (Times, science article, tabloid), comic strips (simplified version of our project), documentary type video?, <br>
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- | Terrorist attacks<br><br>
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- | <b>Characterization presentation by Chris:</b> <br><br> | + | |
- | in vivo presentation<br> | + | |
- | Timer vs. switch. need to know characteristics of parts. <br> | + | |
- | How to characterise<br> | + | |
- | Look at individual parts. What goes in, what goes out. What function is carried out. <br> | + | |
- | What specifically do you want to characterise. <br> | + | |
- | Terminator, core-promoter, RBS, reporter, coding region. <br> | + | |
- | Terminator requires several different characterizations. <br> | + | |
- | Place promoter, coding region into an expression circuit with a reporter. <br> | + | |
- | Standards are only existant for promoters. How strong promoters are. <br> | + | |
- | Canton paper 2008. Base characteristics that you need to know about an inducible promoter. <br> | + | |
- | How responsive (transfer function of response), dynamic? (output) <br> | + | |
- | All relative to J23101. <br> | + | |
- | Synthesis rate, number of cells and divide your promoter output with standard which gives you a value relative to J23101. RPU (relative promoter unit) <br> | + | |
- | RBS calculator gives you a value to how strong it is. Need to debug yourself. <br> | + | |
- | Hard to characterise until we know what we want to do. <br> | + | |
- | Plate readers look at OD, fluorescene, luminescence. <br> | + | |
- | Individual cells analyzed for GFP and RFP. <br> | + | |
- | Chris works on methods to make it more high-throughput (automates to make faster). <br> | + | |
- | Won't consider optimizing something unless it worked twice on the assays. <br> | + | |
- | Fluorescence doesn't give use pinpoint accuracy. Can't be measured really well, only bulk events and not specific events. <br> | + | |
- | Enzymatic activity more done in vitro. <br> | + | |
- | You're working with factories that work depending on what they're in. <br> | + | |
- | Can find Chris though James at any time. <br> | + | |
- | Make sure positive and negative control will work. <br><br> | + | |
- | <b>Antibody research:</b> <br><br>
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- | -Single-domain antibodies such as the nurse shark derived IgNAR and the camelid derived VHH have been used for many purposes and have recently started to gain popularity among the scientific community. <br>
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- | -Contain CD3 loop that gives these Ig's an advantage when looking for cryptic viral epitopes. However, contain ten epitope copies and might still infect. Solved by increasing their mass. <br>
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- | -Both of these Ig's are stable enough to be administered orally. <br>
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- | -Orally administered transformed lactobacilli were used to administer anti-TNF Ig. <br>
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- | -Small dimensions of VHHs allow it to be easily tagged. <br>
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- | -Lactobacilli have been engineered that produces VHH's at a rate fast enough to prevent infection by p2 bacteriophage. Possibly use lactobacillus for screening and E. coli for secretion? <br>
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- | -VHH's and IgNAR's have been effective in detecting poliovirus and inhibiting its replication in vitro, as well as preventing the assembly and secretion of hepatitis B. Possible to use VHH's as intrabodies vs. HIV-1? <br>
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- | -N-glycosylation increases stability. <br>
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- | -Studies demonstrated that pre-immune libraries can be used for rapid generation of Ig's against a large number of harmful antigens. Troublesome low sensitivity overcome by using phage-displayed instead of purified antibodies.[1] <br><br>
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- | <b>References:</b> <br><br>
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- | <a href=http://ultr23.vub.ac.be/ultr/pub/pdfs/98.pdf>[1]</a>Ario de Marco, “Biotechnological applications of recombinant single-domain antibody fragments,” Microbial Cell Factories 10, no. 1 (2011): 44.
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- | </html>
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- | | + | |
- | <html>
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- | <b>Frank Machin</b>
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- | - A method is needed to sort cells by their ability to bind the venom proteins<br>
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- | - One such method would be to have the cells secrete their anti-venom proteins and then flood the cells with venom. The best cells would survive and the weak ones would be killed. However, this may cause the production of proteins that bind the venom components that are less dangerous such as phospholipases and oxidases. It would be more important for the anti-venom to inhibit the neuro-muscular disruptive proteins<br>
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- | <br>
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- | - One effective method would be to use Fluorescence Activated Cell Sorting, which is able to sort cells by their fluorescence. See: http://www.bio.davidson.edu/courses/genomics/method/FACS.html
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- | <br>
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- | - This would require a system that expresses a fluorescent reporter in response to the binding of venom proteins to the cell-surface proteins, but I cannot be too specific until I know how the anti-venom proteins are going to be expressed
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- | <br>
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- | - Yet another method would cause the death of any cells that do not bind to the venom with high enough affinity - it would be similar to the methods employed in T-cell selection in eukaryotic immune systems, but would be more tricky as it has to occur in a bacterium
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- | <br>
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- | - The ideal then, would be to have all the cells that have little or no binding affinity killed, and then those that do bind express GFP so that the fluorescence can be quantified and the binding can be rated
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- | <br>
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- | - Again, I'd need to understand the anti-venom proteins before I could suggest any particular systems<br>
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- | - Perhaps a second species of bacteria could be used to express the venom proteins on their surface, and come into contact with the anti-venom producing cells, causing contact-dependant stimulation, so that those that interact by the venom-antivenom complexes will be stimulated to divide whilst the remaining cells will potentially be killed
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- | See: http://www.ncbi.nlm.nih.gov/pubmed/21085179
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- | </html>
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| | | |
- | <html>
| + | We currently have two running ideas; the anti-venom and a bile acid sensor. We have split off into two teams and each is working hard in order to build a system that the professors might agree to this Friday at 3:30pm. The deadline is approaching and we still require 2 more possible projects to present to the professors.<br><br> |
- | <b>Nikki Kapp</b>
| + | (Text by Rebekka and Chris) |
- | <br>
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- | - we need a method for random mutagenesis of peptides to create high affinity binding proteins to the multiple components of venom<br> | + | |
- | - PCR based mutagenesis is namely used for site directed mutagenesis and has several biases that make it not ideal for random mutation<br>
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- | - in vivo homologous recombination inherent to yeast can be exploited for protein mutagenesis<br>
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- | (Pirakiticulr et al., 2010) | + | |
- | </html>
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