CHARACTERIZATION OF COAT PROTEIN GENE (AV -1) OF Mungbean yellow mosaic virus (MYMV) INFECTING MUNGBEAN IN SOUTH GUJARAT
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Date
2014-09
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PLANT PATHOLOGY DEPT., N.M.COLLEGE OF AGRICULTURE, NAVSARI AGRICULTURAL UNIVERSITY, NAVSARI
Abstract
Aim of the present investigation was to characterize and
use of AV-1 gene of the yellow mosaic disease (YMD) causing
Mungbean yellow mosaic virus for the cloning in the bacterial
system and over expression of the coat protein. The protein over
expressed in bacterial system can be used as antigen to elicit
immune response in animal and production of antibodies for
serological investigation. YMD in Navsari has been observed to
caused by MYMV-Vig:IN:NVS:Mg:2012. The strategies included
isolation of DNA, in silico characterization of the AV-1 gene and its
putative protein for the study of antigenicity and further its over
expression. Disease produce typical yellow mosaic symptoms which
became brighter and enlarge to some extent with the age; however,
depletion of the chlorophyll could not be observed once synthesized.
Severity of the symptoms could be seen in the newly eme rging
leaves. Good quality viral DNA was isolated from younger
symptomatic leaves of mungbean plants by CTAB method and
amplified ~700 bp of the DNA-A molecule of the begomovirus
infecting mungbean. Sequence of MYMV-Vig:IN:NVS:Mg:2012
(JQ004982.1) previously characterized by this laboratory was
retrieved from the NCBI database. The gene was 420 bp long
synthesizing 139 amino acids long putative protein. The same
sequence was used for the study. Sequence was highly conserved.
Epitopes of the amino acids was well spread in the entire amino
acid. Epitopes were found shifted towards the N -terminal site of the
amino acids. Maximum 13 amino acids long peptide sequence was
found in the amino acid sequence starting at 42 positions till 54 t h
positions
(YDNEPSTATVKND)
of
the
putative
amino
acid
sequence. Study of the amino acid sequence indicated that the
hydrophilisity in the sequence was spread all over the sequence;
however, as epitopes, it was also little more shifted towards the N -
terminal region of the protein. The protein was water soluble. Study
of the different properties of protein revealed that the protein being
targeted was of good quality. From the study it was evident that the
putative protein would be of high antigenicity and would produce
good quality antibodies.
Attempt was made to transform the AV -1 gene in the
pETite vector. It could be transformed in the bacteria system,
however, could not over expressed the protein. This might be either
due to the toxic effect of the gene product on the bacterial gr owth
of alteration of essential part of the genome of the bacteria during
the cloning which lead to the lethal effect.
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