Cloning and expression of human growth hormone in E. coli system | ||||
| Arab Journal of Biotechnology | ||||
| Article 3, Volume 18, Issue 1, June 2015, Page 33-44 PDF (521.54 K) | ||||
| Document Type: Original Article | ||||
| DOI: 10.21608/arjb.2015.428408 | ||||
 View on SCiNiTO | ||||
| Abstract | ||||
| The human growth hormone (hGH) stimulates growth, cell reproduction and regeneration in humans. Recombinant hGH (rhGH) is approved for the treatment of multiple human diseases resulting from hGH deficiency. This study aimed to produce a system for production of rhGH using E. coli and pTXB1 expression system. The open reading frame (ORF) of hGH gene includes 573 base pairs was amplified by PCR cDNA clone, using specific primers containing restriction site terminals. The product was cloned into pTXB1 expression vector and transformed into BL21 bacterial cells. The chitin-binding domain (CBD) present in the intein-tag of pTXB1, allowed for the affinity purification of the fusion protein using chitin beads. The molecular weight of the purified protein was determined by SDS-PAGE which revealed a single band at 22 kDa. The purified protein identity was verified using mass spectrometry. Large scale recombinant protein production is becoming increasingly important for applications in the field of proteomics. This study is an initial step for large scale production of purified rhGH for medicinal use as a pharmaceutical product. | ||||
| Keywords | ||||
| Recombinant human growth hormone; protein expression; pTXB1 system | ||||
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