[Cloning and expression in Escherichia coli of oligo-1,6-glucosidase gene from Bacillus subtilis HB002].
The gene coding for oligo-1,6-glucosidase of Bacillus subtilis HB002 was cloned by the shotgun-cloning method and sequenced by the chain-termination method of Sanger et al. It consists of an open reading frame of 1683 bp. The amino acid sequence of oligo-1,6-glucosidase deduced from its nuecleotide sequence predicts a protein of 561 amino acid residues with a Mr of 65.985 kD, which is 81% and 67% identical to those of oligo-1,6-glucosidase from Bacillus sp. and Bacillus coagulans, respectively, 89% and 79% similar to those of oligo-1,6-glucosidase from Bacillus sp. and Bacillus coagulans, respectively. The oligo-1,6-glucosidase gene of Bacillus subtilis HB002 was cloned into Escherichia coli expression plasmid pBV220, the result of SDS-PAGE showed that the oligo-1,6-glucosidase gene had been expressed in Escherichia coli DH5 alpha, the expressed oligo-1, 6-glucosidase has enzymatic activity.
Jiang
Sijing
S
College of Life Science, Hubei University, Wuhan 420062, China.
Ma
Lixin
L
D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology / Q000235:genetics; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009834:Oligo-1,6-Glucosidase / Q000096:biosynthesis / Q000235:genetics*; D016366:Open Reading Frames; D010957:Plasmids; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D009834:Oligo-1,6-Glucosidase