PCR-cloning of the pac gene from E-coli ATCC11105 - Observations on the expression of a recombinant periplasmic protein

被引:0
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作者
Gumusel, F
Kiremit, N
Issever, S
Ogras, TT
Bermek, E
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关键词
penicillin acylase; pac; PCR; cloning; expression;
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中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The penicillin acylase gene (pac) amplified by polymerase chain reaction (PCR) from Escherichia coli (E. coli) ATCC11105 genomic DNA was cloned into pUC9, pEMBL9(+) and pCP40 vectors. A penicillin acylase precursor was overexpressed at 26 degrees C from pac-pEMBL9(+) and pac-pCP40 constructs transformed into E.coli strains JM109 and pCI857 containing HB101, respectively. With the thermo-inducible pac-pCP40 construct some level of mature alpha and beta subunits and varying degrees of enzyme activity were also detected.
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页码:315 / 324
页数:10
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