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Growth promoting activity of Pangasianodon hypophthalmus recombinant growth hormone expressed in Escherichia coli

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Relation http://eprints.cmfri.org.in/10351/
 
Title Growth promoting activity of Pangasianodon hypophthalmus recombinant
growth hormone expressed in Escherichia coli
 
Creator Megarajan, Sekhar
Singh, Shiva Dhar
Angel, Raymond Jani
Meena, D K
Sivakumar, N
Suresh, Easwaran
Kathirvelpandian, A
 
Subject Aquaculture
Fish Biotechnology
Fish Genetics
 
Description Recombinant growth hormone of Pangasianodon hypophthalmus (rPhGH) was efficiently expressed in Escherichia coli
BL 21 (DE3) cells. The expression vector pET-32a(+) was used to clone and express a 550 bp long cDNA fragment, which
encodes the mature region of growth hormone. The rPhGH was expressed as a 6X HIS-tag fusion protein in E. coli upon
induction by Isopropyl b-D-thiogalactoside, and formed insoluble inclusion bodies in the host cells. SDS-PAGE analysis
indicated that the molecular weight of the fusion protein was about 23 kDa, which is comparable to the theoretical value
of the mature growth hormone of the fish. The expressed protein was recovered by solubilising the inclusion bodies under
denaturing conditions with urea and then the denatured proteins were refolded and purified on Ni-NTA column. The purified
recombinant protein was confirmed by Western blot analysis using anti-His antibodies. Total yield of the refolded and
purified protein was 20 mg l-1 of LB medium. Biological activity of the purified recombinant protein was determined in
in vivo bioassay by its ability to promote growth in rohu (Labeo rohita) fingerlings, injected with three different concentrations
of the hormone. A significant increase in growth was observed in rohu fingerlings administered with rPhGH at a dosage of
1.0 mg g body weight -1.
 
Date 2015
 
Type Article
PeerReviewed
 
Format application/pdf
 
Language en
 
Identifier http://eprints.cmfri.org.in/10351/1/12.pdf
Megarajan, Sekhar and Singh, Shiva Dhar and Angel, Raymond Jani and Meena, D K and Sivakumar, N and Suresh, Easwaran and Kathirvelpandian, A (2015) Growth promoting activity of Pangasianodon hypophthalmus recombinant growth hormone expressed in Escherichia coli. Indian Journal of Fisheries, 62 (1). pp. 70-77.