Open AccessMolecular cloning and expression of the xylanase gene from Chainia in Escherichia coli
Author(s) -
Chauthaiwale Vijay M.,
Deshpande Vasanti V.
Publication year - 1992
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1992.tb05579.x
Subject(s) - xylanase , insert (composites) , molecular cloning , restriction enzyme , microbiology and biotechnology , gene , biology , cloning (programming) , genomic library , lac operon , oligonucleotide , ecori , gene expression , biochemistry , peptide sequence , enzyme , materials science , computer science , programming language , composite material
A complete genomic library of Chainia was constructed in coliphage λ vector gt10 and was screened for the xylanase gene using an 18‐mer mixed oligonucleotide probe corresponding to a six‐amino acid sequence of low molecular mass Chainia xylanase. Inserts from 11 putative clones, showing hybridization with the oligonucleotide probe at medium stringency, were subcloned in pUC8 and screened for xylanase gene expression using anti‐xylanase antibodies. The restriction map of the insert (1.4 kb) from one of the four immunopositive clones (PVX8) showing detectable xyalanase activity was constructed. The xylanase activity of PVX8 was not induced by IPTG or xylan. Reorientation of the insert by directional cloning into pUC9 had no effect on the xylanase activity suggesting that an indigenous promoter from Chainia is responsible for the xylanase activity.