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CONSTRUCTION OF PLANT TRANSFORMATION VECTOR: SUB-CLONING OF PIGEONPEA LECTIN (CCL) GENE UNDER rolC PROMOTER

KrishiKosh

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Title CONSTRUCTION OF PLANT TRANSFORMATION VECTOR: SUB-CLONING OF PIGEONPEA LECTIN (CCL) GENE UNDER rolC PROMOTER
M Sc
 
Creator DEEPAK VISHWANATH PAWAR
 
Contributor Rekha Kansal
 
Subject ---
 
Description T-8621
The 928bp rolC promoter (accession number DQ-160187) was PCR amplified
from the bacterial culture of rolC promoter cloned in pUC29 vector using promoter
specific primers containing restriction enzyme sites of XbaI in forward and EcoRI in
reverse. The amplified product was then restricted with restriction enzymes XbaI &
EcoRI and ligated with pORE O4 binary vector digested with the same restriction
enzymes. 828bp Pigeonpea lectin (CCL) was restricted from the bacterial culture of
pigeonpea lectin gene cloned in pGEMT vector with the restriction enzyme NotI and
ligated with the promoter construct such that CCL gene will be cloned downstream of
rolC promoter in pORE O4 binary vector. The clones were confirmed by restriction
digestion and colony PCR, The gene construct was mobilized into Agrobacterium
tumefacience GV-3101, which will be used for transformation of Brassica juncea to
express pigeon pea lectin protein in phloem sap which is desirable in case of
Hemipteran (phloem sap sucking) pests like aphids.
Keywords: aphid, pigeonpea lectin (CCL) gene, rolC pr
 
Date 2016-09-21T16:34:46Z
2016-09-21T16:34:46Z
2012
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/78183
 
Format application/pdf
 
Publisher IARI, NATIONAL RESEARCH CENTRE ON PLANT BIOTECHNOLOGY