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Characterization of cryila and crylle from native bacillus thruingiensis isolates

KrishiKosh

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Title Characterization of cryila and crylle from native bacillus thruingiensis isolates
 
Creator Mary Suchita Xalxo
 
Contributor M.S.Kuruvinashetty
 
Subject Plant Bio-Technology
 
Description Bacillus thuringiensis is a gram-positive spore forming bacterium that is a valuable source
of cry genes that encoding delta endotoxins, activeagainst a wide range of insect pests. In the
present study, an attempt was made to clone and express cry1Ia and cry1Ie from native
Bacillus thuringiensis isolates. Amplicon of 2.1kb obtained on PCR amplification of B.
thuringiensis YESP3 and SW-18 total DNA using cry1Ia and cry1Ie specific primers was
cloned into linear pTZ57R/T. the recombinant clones pSKK2010 and pSKK1401 obtained
were positive for restriction analysis and specific amplification of cry1Ia and cry1Ie. Further,
the cry1Ia and cry1Ie gene was subcloned into prokaryotic expression vectors pET28+ and
expressed in E.coli BL21 (DE3)pLysS. The recombinant clones pSKK2001and pSKK2002
were positive for PCR amplification and restriction analysis. The SDS-PAGE and bioassay
studies showed and expression of 77.7kDaa protein and the protein expressed was toxic to
Plutella xylostella. Sequencing of full-length 2.1kb cry1Ia and cry1Ie amplicon present in
pSKK2010 and pSKK1401 by using M-13 forward and reverse primers reveled that the
cloned gene has 99% homology with reported gene. Further, the cry1Ia and cry1Ie was
subcloned into plant transformation Vector pHS100. The recombinant obtained were named
as pSKK2402 and pSKK2602 were positive for restriction and PCR analysis.
 
Date 2016-10-18T08:49:01Z
2016-10-18T08:49:01Z
2006
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/80754
 
Format application/pdf
 
Publisher UAS, Dharwad