摘要
根据NCBI GenBank上登录的猪IFN-α基因序列设计了 1 对引物,应用 PCR技术直接从3周龄新兴猪肝细胞中扩增出了约 500 bp 的基因片段;将该片段克隆到 pMD 18-T 载体,经PCR、酶切及序列测定,该克隆片段由501个核苷酸组成,共编码 166 个氨基酸,与 NCBI GenBank上登录的 2 个猪 IFN-α基因序列(登录号为 M28623 和 X57191)比较,核苷酸的同源性分别为99.4%和99.2%。
A specific pair of primers was designed and synthesized according to thesequence of swine IFN-α gene published NCBI GenBank, and a fragment of 500bp was amplified by PCR from the liver of (3-weeks-old) swine and the fragment was cloned into pMD18-T vector. The recombinant plasmids were (transferred) into the E.coli DH5α competent cells, which were incubated on ampicillin^(+)plates with X-gal and IPTG at 37℃overnight. The white clones were selected and incubated. The restriction endonuclease (analysis), PCR and sequence analysis indicated that the swine IFN-α cDNA was 501bp in full-length and (encodes) 167 amino acids. The homologies of nucleotide sequence between the swine IFN-α gene and M28623 and X57191 from GenBank were 99.4%and 99.2% respectively.
出处
《中国兽医科技》
CSCD
北大核心
2005年第2期143-146,共4页
Chinese Journal of Veterinary Science and Technology
