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      https://www.ias.ac.in/article/fulltext/jbsc/026/03/0315-0324

    • Keywords

       

      cDNA cloning; growth hormone; expression inE. coli; Heteropneustes fossilis ; Indian catfish; RT-PCR; sequencing; zebrafish

    • Abstract

       

      A tissue-specific cDNA library was constructed using polyA+ RNA from pituitary glands of the Indian catfishHeteropneustes fossilis (Bloch) and a cDNA clone encoding growth hormone (GH) was isolated. Using polymerase chain reaction (PCR) primers representing the conserved regions of fish GH sequences the 3′ region of catfish GH cDNA (540 bp) was cloned by random amplification of cDNA ends and the clone was used as a probe to isolate recombinant phages carrying the full-length cDNA sequence. The full-length cDNA clone is 1132 bp in length, coding for an open reading frame (ORF) of 603 bp; the reading frame encodes a putative polypeptide of 200 amino acids including the signal sequence of 22 amino acids. The 5′ and 3′ untranslated regions of the cDNA are 58 bp and 456 bp long, respectively. The predicted amino acid sequence ofH. fossils GH shared 98% homology with other catfishes. Mature GH protein was efficiently expressed in bacterial and zebrafish systems using appropriate expression vectors. The successful expression of the cloned GH cDNA of catfish confirms the functional viability of the clone.

    • Author Affiliations

       

      Vikas Anathy1 Thayanithy Venugopal1 Ramanathan Koteeswaran1 Thavamani J Pandian1 Sinnakaruppan Mathavan1 2

      1. Department of Genetics, School of Biological Sciences, Madurai Kamaraj University, Madurai - 625 021, India
      2. Genome Institute of Singapore, National University of Singapore, 1, Research Link, IMA Building No. 04-01, Singapore - 117 604
    • Dates

       
  • Journal of Biosciences | News

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