• Inverse polymerase chain reaction mediated chromosome walking within the human glutamic acid decarboxylase gene

    • Fulltext

       

        Click here to view fulltext PDF


      Permanent link:
      https://www.ias.ac.in/article/fulltext/jbsc/023/03/0265-0269

    • Keywords

       

      Introns; polymerase chain reaction; glutamic acid decarboxylase

    • Abstract

       

      Using inverse polymerase chain reaction (PCR), we have cloned partial intronic sequences from human glutamic acid decarboxylase (GAD) gene. A small 153 bp core region was selected from the GAD cDNA sequence to design outward primers corresponding to its 3′ and 5′ ends. EcoRI digested human DNA which had been circularized by self-ligation and then linearized withSacII was used as a substrate to can.y out PCR. This gave a 900 bp long product which was cloned into pUC19. The sequence analysis of this fragment revealed the presence of introns in the region flanking the selected core DNA. In this work we used this technique to walk into the upsteam region of the GAD gene using sequence information from its cloned cDNA.

    • Author Affiliations

       

      Neelima Khairatkar-Joshi1 Nasir S Ahmad1

      1. Department of Natural Product Biology, Research Centre, Hoechst Marion Roussel Ltd., LBS Road, Mulund (W), Mumbai - 400080, India
    • Dates

       
  • Journal of Biosciences | News

© 2017-2019 Indian Academy of Sciences, Bengaluru.