Expression of common carp growth hormone in the yeast Pichia pastoris and growth stimulation of juvenile tilapia (Oreochromis niloticus)
文献类型： 外文期刊
作者： Li, YH 1 ; Bai, JJ 2 ; Jian, Q 2 ; Ye, X 2 ; Lao, HH 2 ; Li, XH 2 ; Luo, JR 2 ; Liang, XF 2 ;
作者机构： 1.Minist Agr PRC, Pearl River Fisheries Res Inst, CAFS Key Lab Trop & Subtrop Fish Breeding & Culti, Guangzhou 510380, Peoples R China
2.Minist Agr PRC, Pearl River Fisheries Res Inst, CAFS Key Lab Trop & Subtrop Fish Breeding & Culti, Guangzhou 510380, Peoples R China; Jinan Univ, Coll Life Sci & Technol, Dept Biotechnol, Guangzhou 510632, Peoples R China
关键词： Common carp (cyprinus carpio);Gh cdna;Pichia pastoris;Secreted;Molecular-cloning;Escherichia-coli;Body-composition;Cdna;Gene
期刊名称：AQUACULTURE （ 影响因子：4.242； 五年影响因子：4.723 ）
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收录情况： SCI
摘要： The cDNA encoding mature growth hormone (GH) was cloned from the pituitary of common carp (Cyprinus carpio) by RT-PCR. To construct the expression plasmid, the GH cDNA was inserted into the pUC18 plasmid and subsequently subcloned into vector pPlCZalphaA, which contains the promotor from the alcohol oxidase (AOX1) gene and the a-factor signal peptide sequence. The yeast Pichia pastoris GS115 strain was transformed with the expression plasmid. Transgene expression was observed after screening of the transformants with Zeocin((TM)). Results showed that, with methanol induction, recombinant carp GH (rcGH) had been expressed and exported into the culture medium. The production peaked at 72 h of induction and the optimal pH for expression was 6.0. The yield was 300-400 mg 1(-1) in shaking-flask fermentation medium, accounting for 34.61% of the total supernatant secreted proteins. rcGH was separated through anion-exchange chromatography. The growth rate of juvenile tilapia (Oreochromis niloticus) injected with purified rcGH, was found to be 24.5% and 53.1% higher than the control at the dose of 0.10 and 1 mug g(-1) body weight week(-1) respectively (P<0.001), while the chemical composition of muscle was not affected significantly by the rcGH treatment. The available production of a large quantity of purified rcGH from this work will strengthen the functional study of rcGH for both theoretical and practical purposes.
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