Team:Nevada/Project
From 2011.igem.org
Strongtruong (Talk | contribs) (→Cyanobacteria) |
Strongtruong (Talk | contribs) (→Task 1) Engineer Synechocystis to overproduce the hexose sugars) |
||
Line 76: | Line 76: | ||
https://static.igem.org/mediawiki/2011/d/d4/UNR_Cyano_Metabolic_Process.jpg | https://static.igem.org/mediawiki/2011/d/d4/UNR_Cyano_Metabolic_Process.jpg | ||
- | + | ''' | |
+ | == Task 1) Engineer Synechocystis to overproduce the hexose sugars<br> == | ||
+ | ''' | ||
Cyanobacteria use photosynthesis to provide energy and carbon skeletons for anabolic processes. During the day excess fixed carbon can be converted to the polysaccharide, glycogen and stored for later use. To engineer Synechocystis to overproduce hexose sugars, we will divert carbon away from the glycogen biosynthetic pathway and towards hexose sugar production. To achieve this goal we will create a null mutation in the gene encoding ADP glucose pyrophosphorylase (AGP). AGP specifically converts glucose to ADP-glucose which is the monomeric precursor to glycogen. Synechocystis AGP knockout mutants have been reported to no longer produce glycogen, but instead accumulate high levels of sucrose (Miao et al (2003) FEMS Letters 218: 71-77). Fortunately, sucrose can easily be converted to the hexose sugars, glucose and fructose, in a reaction catalyzed by the enzyme invertase (INV). Therefore, we will introduce and overexpress the INV gene in the AGP mutant background.<br> | Cyanobacteria use photosynthesis to provide energy and carbon skeletons for anabolic processes. During the day excess fixed carbon can be converted to the polysaccharide, glycogen and stored for later use. To engineer Synechocystis to overproduce hexose sugars, we will divert carbon away from the glycogen biosynthetic pathway and towards hexose sugar production. To achieve this goal we will create a null mutation in the gene encoding ADP glucose pyrophosphorylase (AGP). AGP specifically converts glucose to ADP-glucose which is the monomeric precursor to glycogen. Synechocystis AGP knockout mutants have been reported to no longer produce glycogen, but instead accumulate high levels of sucrose (Miao et al (2003) FEMS Letters 218: 71-77). Fortunately, sucrose can easily be converted to the hexose sugars, glucose and fructose, in a reaction catalyzed by the enzyme invertase (INV). Therefore, we will introduce and overexpress the INV gene in the AGP mutant background.<br> | ||
- | + | ''' | |
+ | == Construct Design == | ||
+ | ''' <br> | ||
We will simultaneously create the AGP mutant and the INV overexpressing line by inserting the INV expression construct into the coding region of the Synechocystis AGP gene. This will be accomplished by creating an AGP gene replacement construct in which the INV overexpression gene cassette and a kanamycin resistance gene cassette are inserted into the coding region of a subcloned AGP gene. This dysfunctional AGP construct will then be transformed into Synechocystis and through homologous recombination, replace the native AGP gene. | We will simultaneously create the AGP mutant and the INV overexpressing line by inserting the INV expression construct into the coding region of the Synechocystis AGP gene. This will be accomplished by creating an AGP gene replacement construct in which the INV overexpression gene cassette and a kanamycin resistance gene cassette are inserted into the coding region of a subcloned AGP gene. This dysfunctional AGP construct will then be transformed into Synechocystis and through homologous recombination, replace the native AGP gene. | ||
Line 119: | Line 123: | ||
''' | ''' | ||
+ | |||
== Task 2) Engineer ''Synechocystis'' to secrete hexose sugar to the medium == | == Task 2) Engineer ''Synechocystis'' to secrete hexose sugar to the medium == | ||
''' | ''' |
Revision as of 17:07, 26 September 2011
.firstHeading {
display: none;
}
/* Wiki Hacks - START */
/* Author: Pieter van Boheemen */
/* Team: TU Delft */
/* Thanks guys - Bill Collins */
/* +1 - Douglas Watson */
#globalWrapper {
background-color: transparent;
padding-bottom:0px;
border: none;
}
#content {
background-color: transparent;
border:none;
padding-bottom:none;
margin-bottom:none;
}
#bodyContent {
border: none;
}
#top-section {
height: 5px;
width: 100%;
margin-top: 0 px;
margin-left: auto;
margin-right: auto;
margin-bottom: 0 !important;
/* margin-bottom: 10px; */
padding:0;
border: none;
background-color: rgba(0,0,0,0.1);
}
#p-logo {
height: 203px;
overflow:hidden;
border:none;
display: none;
}
#p-logo img {
display: none;
}
#search-controls {
display:none;
background: none;
position: absolute;
top: 140px;
right: 260px;
width: 240px;
}
.left-menu {
width: 400px;
display: block;
margin-top: 5px;
float: left;
margin-right: 20px;
border: none;
left: 20px;
}
.left-menu ul {
border: none;
height: 100%
}
.left-menu ul {
line-height: 2em;
}
#menubar.right-menu {
width:300px;
height: 100%;
display:block;
float:left;
margin-top: 5px;
padding-left:50px;
border: none;
right:50px;
}
.right-menu ul {
border: none;
}
.right-menu a {
background-color: transparent;
}
#menubar li a{
color: black;
background-color: transparent;
}
#menubar li a:hover {
color:white;
text-decoration:underline;
background-color: rgba(0,0,0,0.5);
}
#menubar li.selected a {
color:gray;
}
#catlinks {
display: none;
}
#footer-box {
position: relative;
top: 150px;
left: -5px;
background-color: transparent;
border: none;
width: 965px;
margin: 0 auto;
margin-bottom:none;
padding-left:5px;
padding-right:5px;
}
#footer {
border: none;
width: 965px;
margin: 0;
padding: 0;
background-color: transparent;
}
#f-list a {
color: black;
font-size: 10px;
}
#f-list a:hover {
color: #666;
}
#footer ul {
margin: 0;
padding: 0;
}
#footer ul li {
margin-top: 0;
margin-bottom: 0;
margin-left: 10px;
margin-right: 10px;
padding: 0;
}
/* Wiki Hacks - END */