• D J Chattopadhyay

      Articles written in Journal of Biosciences

    • Effect of proteolytic digestion on the function of vesicular stomatitis virus ribonucleoprotein complex

      A K Banerjee J Roy D J Chattopadhyay

      More Details Abstract Fulltext PDF

      The nucleocaPsid Protein (49 Kd) of vesicular stomatitis virus is tightly bound to the genome rendering the latter transcriPtionally comPetent. Controlled digestion with chymotryPsin removed a 12 Kd PePtide from the comPlex. The resulting comPlex failed to serve as temPlate for genome transcriPtionin vitro when the Polymerase comPonents L and NS Proteins were added. A temPlate-associated Protein kinase activity was also lost uPon chymotryPsin treatment. However, the cleaved nucleocaPsid Protein (37 Kd) was still caPable of binding tightly with the genome temPlate and retained the ePitoPe recognized by a monoclonal antibody. These results suggest that the nucleocaPsid Protein Possesses seParate domains that mediate binding to Polymerase comPlex and maintain the structural integrity of the template.

    • Mutagenesis study to ameliorate the bacterial expression of phosphoprotein P of vesicular stomatitis virus

      Susanta Pahari D J Chattopadhyay

      More Details Abstract Fulltext PDF

      The phosphoprotein gene of vesicular stomatitis virus, a Rhabdovirus, has been inserted into bacterial expression plasmids containing the Escherichia coli tac promoter and ribosome binding site (RBS). A low level of expression of the protein was detected. Sequence analysis showed the presence of 15 nucleotides in the spacer region i.e., between the Shine-Dalgarno sequence and ATG. Alteration of the distance and the sequence in the spacer region by oligonucleotide-directed mutagenesis revealed a correlation among the expression levels, accessibility of the RBS and requirement for a minimum spacing of at least 7 nucleotides between the Shine-Dalgarno sequence and ATG for optimal gene expression.

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