Mahatma, LalitSolanki, Hiren P.2017-02-282017-02-282014-09http://krishikosh.egranth.ac.in/handle/1/5810003004Aim 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.ennullThesis