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.     



More information about the Methods mailing list