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Validation of tomato leaf curl virus regions for promoter activity

KrishiKosh

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Title Validation of tomato leaf curl virus regions for promoter activity
 
Creator Bharti.Gunjan
 
Contributor P.U.Krishnaraj
 
Subject Plant Biotechnology
 
Description The present study was conducted to clone and analyze the activity of various regions
of promoters from tomato leaf curl virus in E. coli and yeast (S. cerevisiae) and develop a
plant transformation vector using the promoters of ToLCV. In order to define the minimal
region of PRKT17 and PTOLP promoter necessary for activity, deletions were carried out
from 5’ end of both PRKT17 and PTOLP promoter based on the basic characteristics of
promoters .Taking this as a reference A3, A2, B3, B2 were designed. The promoter fragments
A3, A2, B2 were cloned into prokaryotic promoters vector, pKKLUX (BamHI) having cat
(chloramphenicol acetyl transferase) as reporter gene and transformed into E. coli DH5a..
The clone pGBK111 containing A3 region could grow on the plate upto 250 μg per ml but
pGBK110, pGBK112 containing A2 and B 2 region respectively could grow on the plate
having very low concentration of chloramphenicol (5 – 10 μg/ml). A3 (578 bp), A2 (74 bp),
B3 (174 bp), B2 (122 bp) were cloned into pYESSR + Zeo and transformed into yeast.
Zeocin plate assay for the clones pGBK250, pGBK251, pGBK252, pGBK253 was done by
streaking the clones on the plates having zeocin concentration ranging from 25 to 100 μg per
ml. The pGBK251 and pGBK253 containing A3 and B3 region respectively could grow on
the plate upto 50 μg per ml concentration of zeocin. A1 (611 bp), A2 (74 bp), A3 (578 bp) ,
B1 (286 bp) , B2 ( 112 bp ) , B3 ( 174 bp) were cloned into promoter probe vector pVR37
and pVR44 and transformed to Agrobacterium tumefaciens.
 
Date 2016-11-09T18:14:11Z
2016-11-09T18:14:11Z
2010
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/84766
 
Format application/pdf
 
Publisher UAS, Dharwad