detection system for cloning chitinase gene
Chanpen Wiwat - PYMI
pycww at mucc.mahidol.ac.th
Thu Jan 27 03:30:14 EST 1994
I will clone chitinase gene from Bacillus sp. using BluescriptKS vector
and E.coli DH5 as cloning host. I have tried to detect positive clone by
using agar containing Carboxymethyl- chitin as substrate and stain with
congo red. This method can be used very well in detection chitinase
enzyme but I cannot detect chitinase from E.coli (positive control).
(I have exposed E.coli colony to chloroform vapour for 20 min. before
overlay with agar containing CM-chitin.)
Please give me a protocol, if any one have tried them or any suggestions
for the sensitive detection system.
Thanks.
Chanpen Wiwat
Department of Microbiology
Faculty of Pharmacy, Mahidol University.
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