[Two-codon mutagenesis of alpha-amylase gene of Bacillus amyloliquefaciens].
The oligonucleotide encoding Bam HI recognition site having the structure pCGGGATC had been inserted into the recognition sites MspI of the B. amyloliquefaciens alpha-amylase gene, which was cloned in pTG29B plasmid. The alpha-amylase gene had no BamHI sites before mutagenesis. The set of pNSBamHI plasmids with BamHI site at four different positions was obtained. It was shown that all the mutant alpha-amylases possess different specific activities. One of the mutant proteins possesses reduced thermostability. The mutant alpha-amylases can be used for further experiments on protein-engineering of liquefying-type alpha-amylases.
Smirnova
N A
NA
Sorokin
A V
AV
Laptev
D A
DA
Veĭko
V P
VP
Kozlov
Iu I
IuI
D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D003062:Codon; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific; D004795:Enzyme Stability; D005798:Genes, Bacterial; D006358:Hot Temperature; D009154:Mutation; D010957:Plasmids; D012333:RNA, Messenger; D015183:Restriction Mapping; D000516:alpha-Amylases / Q000235:genetics*
D003062:Codon; D012333:RNA, Messenger; D019003:Deoxyribonuclease HpaII; D015252:Deoxyribonucleases, Type II Site-Specific; D000516:alpha-Amylases
Dvukhkodonnyĭ mutagenez gena alpha-amilazy Bacillus amyloliquefaciens