Cloning,Expression,Characterization and
Application of
Bst DNA Polymerase Large Fragment
HUANG Wei-Hua,ZHAI Feng and HONG Guo-Fan
( Shanghai Institute of Biochemistry,the Chinese Academy of
Sciences,Shanghai 200031,China )
Abstract DNA sequencing using
thermostable DNA polymerase is very valuable in molecular biology. From a
specific strain of Bacillus stearothermophilus,the gene of the Bst DNA
polymerase large fragment which had no 5′-3′ exonuclease activity was cloned
and recombined into an expression vector pYZ23,and the constructed plasmid was
introduced into Escherichia coli JF1125,then the expressed protein was
isolated and purified. The genetic engineered product shows characteristics
similar with the purified natural Bst DNA polymerase large fragment.
It will benefit the wide application of Bst DNA polymerase and the
study of the relationship between the structure and function of Bst
DNA polymerase.
Key Words Bst DNA polymerase；cloning；expression；characteristics；applications
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