Ivawati Mohd Yunus (2013) Expression and purification of recombinant GHRH in E.Coli. Masters thesis, Universiti Malaysia Sabah.
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Abstract
The economics of aquaculture are founded upon capacity utilization of available infrastructure, which in turn depend on high growth rates of the selected organism. Endocrine responses to both environmental and genetic variables ultimately control growth in fish and other vertebrates. Evaluation of these hormonal indices could prove useful in development of growth enhancement in fish. The Grouper species are amongst important commercial aquaculture species in Sabah and regarded as the most expensive fish in the local market. The Growth Hormone Releasing Hormone (GHRH) is a 44-aa polypeptide, and the primary function of GHRH is to stimulate Growth Hormone synthesis and secretion from pituitary somatotrophs via binding to its receptor, GHRH-R. A synthetic gene encoding the fusion protein pe/B-GHRH that was originally discovered in Groupers, consisting of a 144 bp DNA sequence was tested to determine its ability to express in Escherichia coli (BL21). The synthetic gene was ligated to the expression vector pET22b followed by transformation into a non-expression host (EcoliTOP 10) for the purpose of DNA sequence analysis. Expression and protein production was carried out using Escherichia coli (BL21). Induction using O.SmM isopropy/ /3-thiogalactosidase (IPTG) at 37°C resulted in expression of 12 kDa protein suggesting that the protein contains a 6X histidine tag and improper signal peptide cleavage. Further purification was done using size exclusion chromatography method and a fraction containing a single protein was obtained. The expressed protein has potential applications in the aquaculture and ornamental fish industry.
| Item Type: | Thesis (Masters) |
|---|---|
| Keyword: | Growth Hormone Releasing Hormone, Grouper, Recombinant protein expression, Aquaculture biotechnology |
| Subjects: | Q Science > QR Microbiology > QR1-502 Microbiology > QR75-99.5 Bacteria |
| Department: | INSTITUTE > Biotechnology Research Institute (BRI) |
| Depositing User: | DG MASNIAH AHMAD - |
| Date Deposited: | 28 Apr 2025 15:52 |
| Last Modified: | 28 Apr 2025 15:52 |
| URI: | https://eprints.ums.edu.my/id/eprint/43588 |
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