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

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      https://www.ias.ac.in/article/fulltext/jbsc/021/05/0687-0697

    • Keywords

       

      VSVP protein ; overexpression ; parameter optimization ; 5′untranslated region ; mutagenesis

    • Abstract

       

      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.

    • Author Affiliations

       

      Susanta Pahari1 D J Chattopadhyay1

      1. Department of Biochemistry, University of Calcutta, 35, Ballygunge Circular road, Calcutta - 700 019, India
    • Dates

       
  • Journal of Biosciences | News

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