猪Tetherin基因的克隆及序列分析

为研究猪Tetherin基因的功能,应用RT-PCR手段从感染猪繁殖与呼吸障碍综合征病毒的猪肺脏组织扩增得到猪源Tetherin基因,将其克隆到p MD19-T载体并进行测序,采用分子生物学软件将其编码的蛋白质序列与其他来源的Tetherin蛋白序列进行比对分析,采用真核转染的方法研究其在MARC-145细胞中的定位情况。结果显示,克隆得到的猪源Tetherin基因全长534 bp;猪源Tetherin蛋白与牛、绵羊、猫、人、黑猩猩、猫头鹰、猕猴Tetherin蛋白同源性分别为41.5%、44.1%、42.3%、43.7%、45.4%、35.6%、42.1%;猪源Tetherin蛋白为跨膜蛋白,该蛋白定位在MARC-145细胞的细胞膜中。

Cloning and Sequence Analysis of Swine Tetherin Gene

In order to study the function of Tetherin gene,the Tetherin gene was detected in swine lung tissue cells which were infected with porcine reproductive and respiratory syndrome virus by the method of RT-PCR. The obtained fragment was ligated into pMD19-T vector and sequenced. The sequences of Tetherin protein was analyzed and predicted by bioinformatics. And the localization was verified in MARC-145 cells. The result showed that the full-length gene of Tetherin was successfully cloned,which contained 534 base pairs(bp). Sequence analysis showed 41. 5% identity with bovine,44. 1% with sheep,42. 3%with cat, 43. 7% with human, 45. 4% with chimpanzee, 35. 6% with owl, 42. 1% with macaque. Secondary structure prediction and location assay in MARC-145 cells showed that this protein was a transmembrane protein.