猪α-干扰素基因的分子克隆与序列分析

The Molecular Clone and Sequence Analysis of Porcine Alpha Interferon Gene

应用 PCR技术直接从长白猪的肝组织基因组中扩增得到了一条大小约为 50 0 bp的特异带 ,将其连接到 p MD1 8-T质粒中 ,转化感受态的 DH5α,运用蓝白斑筛选、酶切鉴定及质粒 PCR鉴定 ,筛选可疑阳性重组子。测序结果表明 ,该基因 (暂定名为 p IFN-α)由 50 1个核苷酸组成 ,共编码 1 6 6个氨基酸。在核苷酸水平上 ,与 Gene Bank 上序列号为 M2 86 2 3及X571 91的同源性高达 98%和 97.8% ,但存在无义突变 ,氨基酸的同源性达 95.8%。

PCR was applied to amplify a specific segment about 500 bp from liver of Changbai pig directly.It was cloned into the T T windows of plasmid pMD18 T,was transformated into sensetive DH5α.After several identification,suspicious positive recombinant was sequenced,the result demonstrated that it(named pIFN α temporarily) was consisted of 501 nucleotides ,encoding 166 amino acid .Compared with the sequences of M28623 and X57191 in GeneBank,there was 98% and 97.8% nucleotide sequence homologen between them separatelly, but there was insignifical mutation, 95.8% amino acid homologen was deduced between pIFN αand the other two.