Technique / Molecular Biology / Bacterial plasmid bacterial phages vectors
Protein expression in E.Coli
Protein expression in E.Coli
This protocol described a strategy of parallel expression of a protein from a variety of vectors containing different tags and/or fusion partners, and a variety of E. coli host strains.
The strategy to expression recombinant protein in e.coli involves two steps: 1. To Clone your gene of interests into a variety of e.coli expression vectors with different expression tags or fusion proteins and express them in a basic e.coli strain. 2. To clone your gene of interests into a regular e.coli expression vector and express it in a variety of e.coli hosting strains.
Content:
1. Expression in Escherichia coli
2. Optimization of expression levels
3. Improving protein solubility
4. Improving protein stability
5. Decreasing protein toxicity
6. In vitro expression using E. coli extracts
7. Co-expression
Last update 04-Feb-2005, Rating of 10 votes.
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By jaya
on 07-Aug-2008
i was checking the viability of the recombinant E.coli cells from the 50L batch.till the induction with IPTG the viability iam getting is 100%. After 2hrs of induction viability is decreasing to 30%.at 5hrs of induction viability is <10%.I was using the plating method by plaitng the konwn number of cells on the agar plates.cell number was calculated as per the OD. |
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By nancy
on 29-Apr-2008
I have ever met this problem. I try to express NPM-ALK protein in Ecoli. I checked the construct is correct. and I tried different expression condition. Stil I did not get so much protein. Hope to get some help from you. thanks. |
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By alkprotease
on 12-Oct-2007
what is expansion of "DE3" |
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By Vivek Bhalla
on 07-Mar-2007
very helpful, thank you for putting this together; i'm trying to express a large GST-fusion protein and have protein expr at baseline and multiple, shorter protein products with induction. the various suggestions will hopefully work. thank you. |
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By andy
on 28-Mar-2006
hi. |
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By Yen Ngo Thi Hai
on 06-Mar-2006
This is very helpful to me. Thank you very much |
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By xiangwei
on 15-Jan-2006
quite helpful to me. i like it very much |
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By Sonia Avila
on 05-Jan-2006
very good material to increment a research! |
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By Bassam
on 24-Mar-2005
This is a great work and effort! |
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By mikoli oli
on 04-Feb-2005
thx! quite good! |
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