Generation of pcdna 3.1+-gh as a recombinant expression vector of ostrich growth hormone cdna in saccharomyces cerevisiae

摘要

Growth hormone is essential hormone for vertebrates like the ostrich (Struthio camelus) for growth stimulation, carbohydrate metabolism, protein assimilation etc. Growth hormone is secreted by the pituitary gland and expressed in many cells and tissues. The purpose of this study was generation of pcDNA 3.1-GH recombinant expression vector in order to sub-clone ostrich growth hormone cDNA into Escherichia coli. In brief, total RNA was extracted from the pituitary gland tissue and cDNA sample was synthesised. The cDNA was amplified by PCR and revealed a 672 bp fragment on 2% agarose gel electrophoresis. Then, the ostrich growth hormone cDNA was extracted from the gel and was cloned into pCR8/GW/TOPO vector by T/A cloning technique to produce pCR8/GW/TOPO-GH. After obtaining the sequence of cDNA of the ostrich in Iran, it was submitted in GenBank (Accession number: JN559394). Finally, the GH cDNA was sub-cloned using pcDNA 3.1 into Saccharomyces cerevisiae and pcDNA 3.1-GH recombinant expression vector was generated. The results of present study were showed that ostrich growth hormone cDNA was successfully sub-cloned into Saccharomyces cerevisiae. Therefore, the pcDNA 3.1-GH recombinant expression vector generated in this study could be useful to express the ostrich growth hormone in yeast cells as a simple and affordable way to produce this hormone at a large scale.