摘要
本文旨在新疆绵羊生长激素(oGH)基因的克隆。从阿勒泰×中国美利奴杂交羊脑垂体细胞中提取总RNA,分离得到具翻译活性的mRNA。用RTPCR方法扩增出编码oGH的基因,长度为671bp。将扩增产物经琼脂糖凝胶电泳后,回收纯化。采用PCR克隆试剂盒将扩增产物连接至pTAdv质粒上,经PstI和XbaI双酶切鉴定正反插入后,筛选正向插入阳性克隆,并进行序列分析。结果表明克隆到的oGH基因序列与国外报道的序列有4个碱基差异,但所推导的氨基酸序列完全一致。oGH基因的开放阅读框架共含654个核苷酸,编码217个氨基酸,其中信号肽26个氨基酸,编码碱基为1～78(78bp);成熟肽191个氨基酸,编码碱基为79～651(573bp);终止密码子TAG(652～654bp)。其蛋白质的氨基酸序列与牛、人和鱼的序列同源性分别为99.08%、26.7%和5.9%。
To clone and sequence the gene of ovine growth hormone (oGH), the total RNA was extracted from somatotropic cells of pituitary gland of Aletai×Chinese Merino hybrid sheep. The gene of oGH was amplified by RTPCR and its size was about 617 bp. The target segments were separated by agarose gel electrophoresis, excised and purified from the gel,and then cloned into pTAdvs after they had been digested by EcoR I .One positive recombinant clone was sequenced and analyzed .The results showed that the gene of oGH on ORF contains nucleotides 654 bp,coding for 217 amino acids which composed of signalpeptides 26 amino acids and matpeptides 191 amino acids. The homology of amino acid sequence between oGH and cattle, human and fish was 99.08%, 26.37% and 5.9%, respectively. The cloned gene of oGHXJ had 4 nucleotides which were different from the published ones from GeneBank. But the amino acid sequences were the same.
出处
《中国兽药杂志》
2003年第6期8-10,共3页
Chinese Journal of Veterinary Drug
