【摘要】 应用RT PCR技术从草鱼脑垂体总RNA中克隆草鱼生长激素cDNA(cGH),全长669bp,含开放阅读框633bp,编码210个氨基酸,分子量为23.6kDa,等电点为6.28;与已报道的草鱼GH有12个碱基、3个氨基酸残基的差异,同源性为98%.将草鱼cDNA定向插入原核表达载体pGEX 4T 1,构建了重组草鱼GH基因质粒pGEX GcGH,经IPTG诱导,pGEX GcGH在大肠杆菌中可表达49.6kDa的融合蛋白.更多还原

【Abstract】 The cDNA encoding grass carp (Ctenspharynogodom idellus) growth hormone(GcGH) was amplified by reverse transcription polymerase chain reaction(RT-PCR) method using isolated total RNA of grass carp pituitary as the template.GcGH cDNA fragment is 669bp in length,including 633bp of its ORF,encoding 210aa which molecular weight is 23.6 kDa and pI is 6.28.It shares 98% identity to other reported grass carp GH in nucleotide sequence and deduced amino acid sequence.Its expression plasmid pGEX-GcGH was constructed by inserting it into pGEX-4T-1 plasmid and the recombinant plasmid was transformed into E.coli DH5α cells. The expression protein was assayed by SDS-PAGE and a new specific protein band was found with molecular mass of about 50 kDa which is consisted of a 23.6 kDa protein deduced from the GcGH and GST (26 kDa).更多还原