摘要
目的:克隆猪T细胞受体γ链(pTCR-γ)基因,用以研究猪T细胞受体分子结构与功能。方法与结果:以GenBank上登载的pTCR-γ基因为参考序列,用RT-PCR法从猪外周血淋巴细胞中克隆pTCR-γ基因。序列分析表明,pTCR-γ基因开放读框为1026bp,编码341个氨基酸残基,含有由23个氨基酸残基构成的信号肽序列;与参考序列相比,在核苷酸和推导氨基酸序列上的同源性分别为95.6%和88.8%;系统进化分析发现,该基因与绵羊、恒河猴、人、马、牛的同源性相对较高,与褐鼠、家犬、家鼠、家猫的同源性次之,而与禽类、鱼类的同源性最低;生物信息学结构预测表明猪T细胞受体γ链含2个结构域,其中1个为IG_LIKE1结构域(IGv结构域),由第14～114位共101个氨基酸残基组成;另一个为IG_LIKE2结构域(IGc结构域),由第143～238位共96个氨基酸残基组成。结论:克隆并应用生物信息学技术分析了猪T细胞受体γ链基因序列及编码蛋白的结构特征,为进一步研究γ链的结构与功能奠定了基础。
Objective：To clone porcine T cell receptor gamma chain（pTCR-γ） gene for the study on molecular structure and function of porcine T cell receptor.Methods Results：Based on the gene sequence of pTCR-γ from the GenBank database,pTCR-γ gene was cloned by reverse transcription-polymerase chain reaction from porcine peripheral-blood lymphocyte.Sequence analysis showed that pTCR-γ gene was 1026 bp in length and encoded 342 amino acids,including a signal peptide of 23 amino acids.Comparing with the reference sequence,pTCR-γ gene shared 95.6% homology with the published sequence of nucleotide and 88.8% homology of amino acids.The homology was high comparatively with Ovis aries,Macaca mulatta,Homo sapiens,Equus caballus and Bos taurus,second with Rattus norvegicus,Mus musculus,Canis lupus familiais and Felis catus,the lowest with Gallus and Paralichthys.Two domains had been found by the structure prediction of bioinformatics,one was IGv domain that contains 101 amino acids with position of 14～114,another was IGc domain that contains 96 amino acids with position of 143～238.Conclusion：The porcine T cell receptor gamma chain gene was cloned,and its gene sequences and structural feature of encoding protein were analyzed by bioinformatics.This study laid a foundation for further research of structure and function of porcine T cell receptor gamma chain.
出处
《生物技术通讯》
CAS
2010年第2期227-231,共5页
Letters in Biotechnology
基金
国家高技术研究发展计划(2006AA10A203)
国家科技支撑计划(2007BAD40B03)
关键词
猪T细胞受体γ链
基因克隆
生物信息学
结构预测
porcine T cell receptor gamma chain gene cloning bioinformatics structure prediction
