番鸭IFN-α基因的克隆与序列分析

Cloning and Sequence Analysis of Muscovy Duck Interferon-Alpha

采用RT-PCR方法从番鸭肺脏器官中扩增IFN-α基因片段,将扩增产物经胶回收后克隆到pMD18-T载体上,经PCR和双酶切鉴定为阳性后进行序列测定,并用Lasergene v7.1DNAStar软件对测序结果进行分析。结果表明,所扩增的IFN-α基因编码完整的开放阅读框,基因长度为576bp,为国内外首次报道。并与GenBank中登录的各品种鸭、鹅IFN-α基因核苷酸序列进行同源性比较分析,其同源性分别在97.0%～97.7%和95.5%～97.7%;遗传进化分析表明,鸭和鹅IFN-α基因在遗传进化上呈各自独立的分支,推测IFN-α基因在不同动物中有严格的种属特异性。

Reverse transcription-polymerase chain reaction（RT-PCR） method was used to amplified the IFN-α gene fragment from lung tissue of Muscovy duck,then cloned to pMD18-T vector,the positive clones were confirmed by PCR and restriction enzyme digestion,the obtained sequence were analyzed by the Lasergene v7.1 DNAStar software.The results showed that,the Muscovy duck IFN-α gene encoding a 576 bp open reading frame（ORF）,which was first reported in the world.The nucleotide had higher similarity ranging from 95.5% to 97.7% among different goose species,also had higher similarity ranging from 97.0% to 97.7% among different duck species;however,the ducks and geese IFN-α gene had its＇ own phylogentic lineage,which means,the IFN-α gene had strictly specificity in different species.