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Technique / Proteomics and protein biochemistry / Protein expression, protein extraction and protein purification / E.coli expression system
| Protocols for protein expression in e.coli host including expression vector and host strain selection, induction condition optimization, tagged protein purification etc. |
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(E.coli protein expression reviews) 1: Peti W, Page R. Strategies to maximize heterologous protein expression in Escherichia coli with minimal cost. Protein Expr Purif. 2007 Jan;51(1):1-10. Epub 2006 Jul 4. Review. 2: Hart DJ, Tarendeau F. Combinatorial library approaches for improving soluble protein expression in Escherichia coli. Acta Crystallogr D Biol Crystallogr. 2006 Jan;62(Pt 1):19-26. Epub 2005 Dec 14. Review. 3: Sorensen HP, Mortensen KK. Advanced genetic strategies for recombinant protein expression in Escherichia coli. J Biotechnol. 2005 Jan 26;115(2):113-28. Review. 4: Baneyx F. Recombinant protein expression in Escherichia coli. Curr Opin Biotechnol. 1999 Oct;10(5):411-21. Review. 5: Yee L, Blanch HW. Recombinant protein expression in high cell density fed-batch cultures of Escherichia coli. Biotechnology (N Y). 1992 Dec;10(12):1550-6. Review. 
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Last update: 
17-May-2008 03:18 am
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An E. coli expression system optimized for DELLA proteins. Protein Expr Purif. 2008 Mar;58(1):168-74
Authors: Sun X, Frearson N, Kirk C, Jones WT, Harvey D, Rakonjac J, Foster T, Al-Samarrai T
The DELLA proteins are involved in regulation of plant growth in response to phytohormonal signals such as GA, ethylene, and auxin. They have become one of most challenging and active area of research due to their fundamental roles in plant biology. Here, we describe the first successful expression of the N-terminal domains of DELLA proteins of Arabidopsis thaliana and Malus domestica in Escherichia coli system which will be used to produce monoclonal antibodies for profiling protein micro-arrays. Optimizations of the cloning, expression, and purification using specific tags have been discussed.
Molecular cloning, expression, purification, and characterization of shorter forms of human glutamic decarboxylase 67 in an E. coli expression system. Brain Res Mol Brain Res. 2005 May 20;136(1-2):255-61
Authors: Sha D, Wei J, Wu H, Jin Y, Wu JY
Previously, we reported the presence of truncated form of human brain l-glutamic decarboxylase 65 (tGAD65) in vivo as well as in vitro and found that tGAD65 was more active than the full-length GAD65 (Wei et al., J. Biomed. Sci., 10: 617-624, 2003). Here, we report the presence of two shorter forms of hGAD67, namely, hGAD67 (Delta1-70) and hGAD(67) (Delta1-90), referring to a deletion of 1-70 and 1-90 amino acids from the N-terminal, respectively. The molecular masses of hGAD67 (Delta1-70) and hGAD67 (Delta1-90) were found to be 59 kDa and 57 kDa, respectively. Both shorter forms were cloned, expressed, and characterized. In contrast to hGAD65, the shorter forms of hGAD67 were much less active than the full-length due to decrease in affinity of PLP towards the shorter enzymes. Both the full-length and one of the shorter forms of GAD67 were detected in porcine brain extract. Furthermore, the full-length GAD67 could be converted to both shorter forms by crude brain extract, suggesting that an endogenous protease may be present in the brain, which is responsible for the conversion. The cleavage of GAD67 seems to be Ca+(2)-dependent. The model for the conversion of GAD from full-length GAD to shorter forms of GAD and its physiological implications was proposed.
E. coli expression system for identifying folding mutations of human adenosine deaminase. Methods Mol Biol. 2003;232:175-82
Authors: Santisteban I, Arredondo-Vega FX, Daniels S, Hershfield MS
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