Journal of Marine Science and Technology

Journal of Marine Science and Technology

Molecular cloning and sequence analysis of growth hormone cDNA of Orange-spotted grouper (Epinephelus coioides) in E.coli

Document Type : Original Manuscript

Authors
f kh 1
Hossein Zolgharnien 2
Bita Archangi 2
Ebrahim Rajabzadeh 3
Ahmad Qasemi 4
1 1 Department of Marine Biology, Faculty of Marine Science and Technology, University, Khorramshahr, Iran
2 Department of Marine Biology, Faculty of Marine Sciences and Oceanography, Khorramshahr University of Marine Science and Technology
3 Department of Fisheries, Faculty of Marine Natural Resources, Khorramshahr University of Marine Science and Technology
4 Studies and Research Center of Persian Gulf
10.22113/jmst.2016.8234
Abstract
According to the importance of growth hormone gene in aquaculture, in this study, cloning of gowth hormone gene of Orange-spotted grouper (Epinephelus coioides) was examined in pTZ57R/T vector.For this purpose, growth hormone cDNA was preparated and amplified using specific designed primers. PCR product was purified by QIAquick Gel Extraction Kit, Growth hormone gene was inserted in pTZ57R/T vector. Ligation product was transformed into competent E.coli (DH5α) cells. The white clones selected on the LB media and were extracted plasmid as recombinant bacteria .verified clones obtained in this study were performed by direct PCR and sequencing methods Orange-spotted grouper growth hormone cDNA contains an open reading frame consisting of 615 nucleotides and 204 amino acid protein. The calculated molecular weight and isoelectric point of growth hormone protein were respectively 23.014 and 6.9. Results of this study suggested that growth hormone gene of Orange-spotted grouper was successfully cloned in the pTZ57R/T vector, and can be used for growth hormone gene expression in expression vectors and growth hormone protein production. Comparison of gene sequences obtained with E.coioides GH gene sequence in gene bank, indicate sequence similarity between them.
Keywords
growth hormone
Cloning
Epinephelus coioides
pTZ57R/T vector

Subjects

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Journal of Marine Science and Technology
Volume 14, Issue 4
Winter 2016
Pages 108-117

  • Receive Date 19 November 2013
  • Revise Date 23 December 2014
  • Accept Date 24 February 2015
  • Publish Date 20 February 2016