鲤鱼生长激素基因的改造及其在大肠杆菌中的表达

Modification of Carp Growth Hormone Gene and its Expression in Escherichia coli

用聚合酶链式反应（ＰＣＲ）改造了鲤鱼生长激素基因，扩增出编码鲤鱼生长激素成熟肽的序列，并将此序列克隆到ｐＢｌｕｅｓｃｒｉｐｔⅡＫＳ质粒中，序列分析后，亚克隆到大肠杆菌表达载体ＰＢＶ（２２０）中。经ＳＤＳ—ＰＡＧＥ和薄层扫描，结果表明鲤鱼生长激素基因已在大肠杆菌中获得表达。鲤鱼生长激素成熟肽的分子量为２２０００道尔顿，表达率占细胞总蛋白的１８％以上。

Carp growth hormone gene is modified by using the Polymerase Chain Reaction (PCR),the plasmid which contains carp growth hormone genes is used to amplify mature polypeptidesequence of encoding carp growth hormone. PCR products are cloded into plasmid pBluescript Ⅱ KS+/- to make DNA sequencing, then subcloned into expression plasmid PBV220 inEscherichia coli. The results of SDS-PAGE and scanning oensitometer show carp growthhormone has obtained expression in Escherichia coli. Molecuiar weight of carp growth hormone is 22000 dations, the total yields of carp growth hormone occupy over 18% of total cellular protein.