苏钟猪ACE2基因克隆及其遗传进化分析

Cloning of ACE2 gene of Suzhong pigs and its phylogenetic analysis

为了克隆苏钟猪的血管紧张素转化酶2(ACE2)基因,根据GenBank发表的ACE2基因序列,设计合成1对引物,从苏钟猪心脏提取总RNA,对ACE2基因进行RT-PCR扩增并测序,然后利用DNAstar软件对序列进行比对分析。产物经琼脂糖电泳分析,呈现1条约641 bp的条带,回收纯化后对扩增的猪心肌ACE2 mRNA进行了测序和比较分析。扩增的猪ACE2核苷酸序列与GenBank中已登录的2只猪ACE2核苷酸序列同源性分别为98.6%和99.2%。同时,利用DNAstar将其与人、牛、大鼠和小鼠等物种相应序列进行比较,结果表明苏钟猪ACE2基因与牛同源性最高,达到87.5%;与人的同源性为82.5%,与斑马鱼的同源性最低,仅为52.3%。对由苏钟猪ACE2基因核苷酸序列推导的氨基酸序列进行同源性比较也得到了同样的结果。

To clone angiotensin-converting enzyme 2（ACE2） gene of Suzhong pig,one pair of primers was designed based on the ACE2 gene sequences published in GenBank,and the total RNA of ACE2 gene total RNA extracted from Suzhong pig heart was amplified by RT-PCR.The product of 641-bp fragment in length was obtained as expected after agarose gel electrophoresis.The results demonstrated that nucleotide homologies of ACE2 between Suzhong pig and other two pigs published in GenBank were 98.6% and 99.2%,respectively.The nucleotide homology of ACE2 between Suzhong pig and Bos Taurus was the highest,87.5%,by DNAstar,82.5% with Homo sapiens,and 52.3%,the lowest with Danio rerio.The homologous analysis of its amino acids in different species appeared the same results as nuclectide.