Cloning and complete sequence of the DNA polymerase-encoding gene (BstpolI) and characterisation of the Klenow-like fragment from Bacillus stearothermophilus.
A fragment of the DNA polymerase I-encoding gene (polI) from Bacillus stearothermophilus (Bst) was obtained by PCR. This was used as a probe to obtain a full-length gene from a Bst genomic DNA (gDNA) plasmid library. Comparison of the sequence to B. caldotenax (Bca) showed about 93% homology at the amino acid (aa) level. A Klenow-like (BstpolIk) clone was developed and the recombinant protein displayed DNA polymerase activity similar to the wild-type BstPolI enzyme.
Phang
S M
SM
Dept. of Zoology, National University of Singapore.
Teo
C Y
CY
Lo
E
E
Wong
V W
VW
D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004256:DNA Polymerase I / Q000096:biosynthesis / Q000235:genetics*; D004257:DNA Polymerase II / Q000096:biosynthesis / Q000235:genetics; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D015698:Genomic Library; D001411:Geobacillus stearothermophilus / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid
D004256:DNA Polymerase I; D004257:DNA Polymerase II