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  • Cloning and expression of alkaline protease genes from two salt-tolerant alkaliphilic actinomycetes in E. coli.

Cloning and expression of alkaline protease genes from two salt-tolerant alkaliphilic actinomycetes in E. coli.

International journal of biological macromolecules (2012-02-14)
S D Gohel, S P Singh
摘要

Cloning and expression of recombinant alkaline serine proteases from two salt tolerant alkaliphilic actinomycetes strains: OM-6 (EU710555.1) and OK-5 (HM560975) were successfully obtained in mesophilic host, Escherichia coli. The positive clones harboring protease genes were selected on the basis of restriction analysis on agarose gel. The effect of temperature and IPTG concentrations on the expression of recombinant proteases and solubilization of the expressed enzymes was assessed. SDS-PAGE revealed protease bands corresponding to 25 kDa and 20 kDa molecular mass representing OM-6 and OK-5 proteases, respectively. Cloning and expression of alkaline proteases from salt tolerant alkaliphilic actinomycetes would pave the way for further biochemical and molecular characterization to achieve unexplored features of the biocatalysts from extremophiles.

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Sigma-Aldrich
IPTG, ≥99% (TLC), ≤0.1% Dioxane
Sigma-Aldrich
异丙基β-D-1-硫代吡喃半乳糖苷, ≥99% (TLC)
Sigma-Aldrich
异丙基 β-D-硫代半乳糖吡喃糖苷 溶液, ReadyMade IPTG solution for Blue-white screening