CLONING AND PROKARYOTIC EXPRESSION OF GROWTH HORMONE GENE IN SINIPERCA CHUATSI
鳜(Siniperca chuatsi)生长激素基因克隆和原核表达

GH cDNA was amplified and cloned from total RNA isolated from pituitary gland of Siniperca chuatsi by RT-PCR. Sequence analysis showed an open reading frame (ORF) of S. chuatsi growth hormone gene in size of 615bp, encoding 204aa of 23kDa and pI at 7.07, comprising 10.78% acidic, 12.74% basic, 44.12% hydrophobic, and 32.35% hydrophilic. The cDNA of mature growth hormone was directionally cloned in pET-32a(+), and then the recombinant expression vector was transformed into E. coli BL21 (DE3 plys). After induction by IPTG, an expected 43kDa fusion protein was found by SDS PAGE analysis, accounting for 58% of the total protein. Western blot demonstrated that the recombinant fusion protein can be recognized by anti-6×His monoclonal antibody.