Sequencing of partial cypD gene for screening of Bifunctional Cytochrome P450 Monooxygenases from Bacillus cereus group

Order of Publishing in Issue: 
3
Volume :4
Issue :3
July, 2010
Page No: 
755-763
Authors: 
P. Brahmaiah, T. Sathish, M. Hymavathi, K.Sreedhar Reddy and R. S. Prakasham*
Address: 
Bioengineering and Environmental Centre Indian Institute of Chemical Technology Hyderabad – 500 607, India.
Email-ID: 
prakasam@iict.res.in

Abstract:

Four different PCR primers were specifically designed to amplify a fragment of cypD gene from isolated Bacillus cereus group strains. Amplification was carried out using these primers. Among all isolates, strains designated as CYPPB-1, 2 and 3 possess cypD gene. Phylogenetic and comparative sequence analysis revealed differences among the sequences of isolated B .cereus group strains as well as those from the public databases. Sequences of all three B. cereus group isolates clustered with that of B. cereus G9842 and there is high similarity between sequences of CYPPB-1 and 3 where as CYPPB-2 showed a little variation. Further microscopic and 16S rDNA analysis indicated that CYPPB-1, 2 belongs to B.cereus while CYPPB-3 to B.thuringiensis. Comparative translated aminoacid sequence analysis with respect to B.cereus 14579 revealed that serine instead of proline at position 570 in all three isolated strains and histidine instead of glutamine at position 602 in CYPPB-1 and 3 strains. This diversity of CYP102A in B. cereus group may signify potential resource of different bifunctional cytochrome P450 monooxygenases.

Keywords: 
cypD gene, Bifunctional Cytochrome P450 Monooxygenase, B.cereus group, primer design, amplification and sequence analysis.
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