鲤鱼生长激素基因的PCR扩增、克隆及其序列比较  

Amplificaton, Cloning and Sequence Comparison of Growth Hormone Gene for Carp by Polymerase Chain Reaction

作　　者：赵晓祥[1] 李晶[1] 李荣萍[1] 张淑梅[1] 杨志兴[1] 

机构地区：[1]黑龙江省科学院应用微生物研究所,哈尔滨150010

出　　处：《生物工程学报》1996年第3期284-288,共5页Chinese Journal of Biotechnology

基　　金：黑龙江省"八五"攻关资助项目

摘　　要：从鲤鱼(Cyprinus carpio)脑垂体中分离其总RNA,经反转录成为第一条链cDNA。以第一条链cDNA为模板,以合成的两段29个寡聚核苷酸为引物,再经过PCR扩增,合成了鲤鱼的生长激素基因,并把其克隆到大肠杆菌表达载体(p^(Blue-scriptⅡ KS+/-))上。序列分析和限制酶切图谱表明所克隆的鲤鱼生长激素基因的开放读框含有630bp,编码210个氨基酸,其中含有22个氨基酸的信号肽和188个氨基酸的成熟GH序列。我们所克隆的鲤鱼GH基因与Koren发表的鲤鱼GHcDNA的序列在编码区完全一致,但是与Chao发表的鲤鱼GHcDNA比较,在编码区核酸序列和氨基酸序列的同源性分别为95.6％和96.7％。Total RNA was isolated from carp (Cyprinus carpio) pituitary gland. Then the first strand cDNA was synthesized using oligo (dT) 12-18 as primer, total RNA as template and AMV reverse transcriptase. Next the polymerase chain reaction (PCR) was performed using the first strand cDNA as template, synthetic 29 oligonucleotides as primer and Taq DNA polymerase (94C60s, 55C30s, 72C50s, 35 cycles). After PCR amplification, the products were cloned into an E. coli expression vector ( pBluesctipu KS+/-). The result of sequence analysis and restriction map shows that an open reading frame of carp growth hormone gene contains 630 base pairs which code for a polypeptide of 210 amino acids including 22 amino acids of the signal peptide and 188 amino acids of the mature growth hormone. Nucleotide sequence and amino acid sequence of carp growth hormone gene are as same as Keren's carp GH cDNA in code region. Compared with Chao's carp GH cDNA.homology of nucleotide sequence and amino acid sequence of carp growth hormone gene is 95. 6% and 96. 7% respectively in code region.

关 键 词：鲤鱼 生长激素基因 聚合酶链反应 克隆 序列分析 

分 类 号：Q959.468[生物学—动物学]