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Studies on identifying efficient PTGS technology in silencing ricin gene in the model plant, tobacco

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Title Studies on identifying efficient PTGS technology in silencing ricin gene in the model plant, tobacco
 
Creator PASULA SOMA SEKHAR REDDY
 
Contributor Sivaramakrishnan, S
 
Subject Studies, identifying, efficient, PTGS, technology, silencing, ricin, gene, model, plant, tobacco
 
Description The present investigation was carried out with an objective of establishing a
model system in tobacco for assessing the efficiency of different PTGS vectors
developed for reducing ricin and RCA in castor. The strategy involved developing
transgenic tobacco expressing full-length preproricin gene that could serve as an
excellent model system to assess the efficiency of the PTGS constructs for silencing
ricin and RCA.
Towards this end, tobacco was transformed, through Agrobacteriummediated
leaf-disc transformation, with a gene cassette containing full-length ricin
gene under CaMV 35S promoter. The transgenicity of the putative transgenics was
confirmed subjecting them to gene specific PCRs. Different combinations of
primers for amplifying various component sequences were used to ascertain that the
presence of the entire gene cassette so that the introduced ricin gene expressed and
produced the transcript. The expression of ricin gene in these transgenics was
confirmed by RT-PCR studies. Again, by using different primer combinations it was
confirmed that the entire transcript of ricin was produced.
The confirmed tobacco transgenics were then re-transformed with the
developed PTGS constructs and the putative transgenic shoots for each of the PTGS
constructs were obtained. However, as the shoots were small and were in clusters,
the molecular analysis could not be completed with these shoots.
Parallel to this, transgenic tobacco plants were developed independently with
the five PTGS constructs. These plants were developed with the objective of
crossing them with the plants expressing ricin gene so that in the progeny the
efficiency of each of the PTGS vectors in silencing of the ricin gene. Transgenic
tobacco harbouring PTGS constructs was confirmed by PCR.
Analysis of the expression pattern and levels of transgenic tobacco
expressing individual PTGS constructs and confirmation of transgenic tobacco retransformed
with individual PTGS constructs would be the future line of work.
 
Date 2016-08-17T13:32:12Z
2016-08-17T13:32:12Z
2006
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/72742
 
Language en
 
Relation D7871;
 
Format application/pdf
 
Publisher ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, RAJENDRANAGAR, HYDERABAD