"

From 2011.igem.org

Monday

Today we laid out the weekly schedule for assembly, as follows

Day 1: grow culture

Day 2: Assemble (all day)

Day 3: grow colonies and check with PCR

Day 4: miniprep

Day 5: sequence results

Day 6: use miniprep

Note: Every step is done each day for different assemblies

We also began developing preliminary ideas for the summer project:

Proposed Idea	Preliminary Thoughts
Bacterial photosynthesis	Up-regulate bacterial chlorophyll, BCHM=gene to increase chlorophyll production
Microbial fuel cells	Bacteria add electrons to Fe+3 and electrodes to create current, Use sacrificial anode (geobacter)
Serotonin sensor	????
Bacterial arithmetic	Use sRNAs to regulate translation to mirror arithmetic operations based on expression rates

We spent the rest of the day researching these ideas to determine their plausibility.

Tuesday

• Jim started the day off with some information regarding geobacter. Jim explained the goal to optimize geobacter’s production of current in the presence of iron. The proteins responsible in geobacter are called cytochromes. In 2010, Missouri University of Science and Technology created a construct containing some of the cytochromes, so Jim proposed that we look into their part and see if we can optimize it further. The problem with working with Geobacter is its slow growth time, which makes growing cultures difficult. Thus, we determined that this project idea would not be suitable for a summer venture.

• Another idea was proposed today regarding radiation sensing and lambda phage repressors. This idea was proposed by Penn State’s 2007 team, but was never really brought to practice. We proposed a device that contained different strains of bacteria that correspond to different amounts of radiation exposure. Each strain would produce a pigment when exposed to a certain threshold of radiation. After receiving unanimous acceptance, we decided to further develop this radiation sensor. We spent the rest of the day researching the lambda phage system and mechanisms involved in radiation exposure.

• One problem that arose was RecA’s constitutive presence in the cell. RecA promotes homologous recombination, but we do not want recombination between our constructed plasmid and the bacteria’s chromosome. Thus, we needed a mutant of RecA that would still work in the lambda phage system without catalyzing homologous recombination.

Wednesday

• Today, many of the students new to iGEM and lab research at Penn State (Vishal, Alex, Jamie, Kristen, Byron) attended a lecture on handling chemicals and hazardous materials given by Penn State’s Environmental Health and Safety department. After reconvening with the rest of the team, we continued discussing our RecA problem. Jim proposed using a different system (phi80) of repressors used to sense DNA damage, but the RecA mutant used in this system had the same problems of lambda phage. We determined that we would have to make our own mutant of RecA and developed a system to test it, as shown in Figure 1.

• We also looked into our reporter system, which would be activated by the lambda system. We researched the iGEM project submitted by Imperial College in 2010 in order to have a fast reporter system for our biological dosimeter.

Back to Notebook

Home	Team	Penn State Official Team Profile	Project	Parts Submitted to the Registry	Modeling	Notebook	Safety	Attributions