علوم زیستی دریا
f kh; Hossein Zolgharnien; Bita Archangi; Ebrahim Rajabzadeh; Ahmad Qasemi
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 ...
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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.