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Construction of synthetic gene encoding for cecropin peptide with broad spectrum antimicrobial activity

KrishiKosh

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Title Construction of synthetic gene encoding for cecropin peptide with broad spectrum antimicrobial activity
 
Creator Manjunath G.Maloji
 
Contributor M.S.Kuruvinashetty
 
Subject Plant Bio-Technology
 
Description In this study, efforts were made to clone synthetic genes cer and msr encoding
proteins having broad-spectrum antimicrobial activity into a prokaryotic expression vector,
plant transformation vector and transformation of cotton. Antimicrobial peptide genes cer and
msr, available in public data bases were codon optimized to suit cotton and custom
synthesized. The codon-optimized genes were cloned in a pET28 prokaryotic expression
vector and E. coli BL-21(plysS) was transformed. SDS-PAGE analysis of the recombinant
clones of E. coli BL-21(plysS), showed the expression of the corresponding protein. The
crude protein extract from gene in pET28a+ having cer and msr were tested against certain
plant pathogenic fungi. The protein caused reduction in the growth of Sclerotium rolfsi,
Rhizoctonia bataticola and Fusarium solani compared to the control. The cer and msr inserts
were further subcloned into a plant transformation vector pHS100 to facilitate plant
transformation. The recombinant clones were mobilized into Agrobacterium tumefaciens
LBA4404 by triparental mating. The A. tumefaciens with recombinant clones were used to
transform cotton. The survived plants on rooting medium with selection (kan 100 μg/ml)
were considered as putative transformants. These plants were further confirmed through PCR
with specific primer.
 
Date 2016-10-18T08:47:34Z
2016-10-18T08:47:34Z
2006
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/80753
 
Format application/pdf
 
Publisher UAS, Dharwad