IGEM:Caltech/2008/Project/Lactose intolerance: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
 
(66 intermediate revisions by 3 users not shown)
Line 1: Line 1:
=Lactose Summary=
<div id="mainpage">
<hr width="750px">
<div id="about">
{{Caltech_iGEM_08}}
</div>
<hr width="750px">
{|cellspacing="5" cellpadding="10" style="background:#ffffff; width: 750px;"
|-valign="top"
|style="background:#ffffff"|
 
==Lactose Summary==
We want to make two plasmids:<br>
We want to make two plasmids:<br>
Plasmid #1 - A constitutive promoter with <i>LacZ</i> and <i>LacY</i>.<br>
Plasmid #1 - A constitutive promoter with <i>LacZ</i> and <i>LacY</i>.<br>
Plasmid #2 - A lactose inducible promoter with lysozyme and holin.<br>
Plasmid #2 - A lactose inducible promoter with lysozyme and holin.<br>
The general idea is to overexpress LacY (Lactose receptor protein) with LacZ (Beta-Galactosidase). When lactose is present, the cell will uptake lactose and will flip on the LacI / LacIQ promoter when enough lactose is present. We then express lysozyme and holin which will lyse the cell releasing beta-gal cleaving lactose into glucose and galactose. Hopefully, glucose and galactose will be taken up by the host in the colon.
The general idea is to overexpress LacY (Lactose transporter) with LacZ (Beta-Galactosidase). When lactose is present, the cell will uptake lactose and will flip on the Lac promoter repressed by lacI when enough lactose is present. We then express lysozyme and holin which will lyse the cell releasing beta-gal cleaving lactose into glucose and galactose. Glucose and galactose will then be taken up by the host in the colon.<br>
==[[/Lactose Outline|Lactose Outline]]==
 
 
 
 
 


==Lactose Outline==
|} </div>
<b>6.19.08</b><br>
We have decided to use the I732017 LacZ (1009/11H) because we could not transform I732005 LacZ (1018/6F).<br>
We ordered primers for LacY, and we will need to PCR out LacY. We then need to run cite directed mutagensis on LacY by added 2H to prevent EIIA(GLUC) binding.<br>

Latest revision as of 15:52, 15 August 2008

iGEM 2008

Home        People        Project        Protocols        Notes        Ideas        Changes       

Lactose Summary

We want to make two plasmids:
Plasmid #1 - A constitutive promoter with LacZ and LacY.
Plasmid #2 - A lactose inducible promoter with lysozyme and holin.
The general idea is to overexpress LacY (Lactose transporter) with LacZ (Beta-Galactosidase). When lactose is present, the cell will uptake lactose and will flip on the Lac promoter repressed by lacI when enough lactose is present. We then express lysozyme and holin which will lyse the cell releasing beta-gal cleaving lactose into glucose and galactose. Glucose and galactose will then be taken up by the host in the colon.

Lactose Outline

Retrieved from "https://openwetware.org/mediawiki/index.php?title=IGEM:Caltech/2008/Project/Lactose_intolerance&oldid=231782"