陆川猪FAM134B基因克隆及生物信息学分析

Cloning and Bioinformatics Analysis of FAM134B Gene in Luchuan Pig

本试验旨在对陆川猪FAM134B基因进行克隆及生物信息学分析。本研究利用Gen Bank公布的猪序列设计引物,应用RT-PCR扩增得到目的基因片段,应用生物信息学方法分析和预测了陆川猪FAM134B基因的理化性质与二级结构。结果显示:陆川猪FAM134B基因编码区全长1 107 bp,编码368个氨基酸;陆川猪FAM134B基因与与金华猪(JX854456.1)、牛(KM587693.1)、山羊(KF684947.1)、人(NM_001034850.2)、小鼠(NM_001034851.2)、大鼠(NM_001034912.1)、非洲爪蟾(NM_001126975.1)序列相似性分别为87.80%、87.44%、86.9%、68.47%、67.01%、67.29%、61.44%,结合系统进化树分析,结果表明不同物种FAM134B基因在进化过程中具有高度保守性。通过比较分析发现,陆川猪FAM134B基因CDS序列区比NCBI公布的金华猪FAM134B序列(JX854456.1)多了一段129 bp序列,比预测的猪FAM134B序列(XM_003483804.2)少了一段371 bp,这提示所扩增得到的陆川猪FAM134B基因有可能是另外一种剪接体。本研究成功克隆陆川猪FAM134B基因完整的CDS序列,为今后FAM134B基因在陆川猪的脂肪沉积及脂肪代谢方面的研究提供基础理论。

The objective of this study was to clone the FAM134 B gene of Luchuan pig and to analyse its genetic structure with bioinformatics. A pair of special primers was designed according to predicted sequence of pig FAM134 B gene in Gen Bank. The FAM134 B gene was amplified by RT-PCR and the physicochemical property and secondary structure of FAM134 B gene were systemically analyzed by bioinformatics techniques. The results showed that the cloned FAM134 B gene fragment included an 1 107 bp whole length CDS（coding 368 bp aminoacids）. The sequence multi-aligned results showed that Luchuan pig shared 87.80%,87.44%,86.9%,68.47%,67.01%,67.29% and 61.44% of similar nucleotide sequence with that of Jinhua pig（JX854456.1）,Bos（KM587693.1）,Capra hircus（KF684947.1）,Homos（NM_001034850.2）,Mus（NM_001034851.2）,Rattus（NM_001034912.1） and Xenopus（NM_001126975.1）,respectively. Comparing with the sequence of Jinhua pig,the CDS sequence of FAM134 B gene was inserted a fragment of 129 bp in Luchuan pig. But the CDS sequence of FAM134 B gene in Luchuan pig was deleted a fragment of 371 bp according to the predicted sequence of pig FAM134 B gene in Genbank. These results indicated that it might be other spliceosome of FAM134 B gene in Luchuan pig. Our study suggested that the whole CDS sequence of FAM134 B gene was successfully cloned in Luchuan pig. And the cloning and analysis of FAM134 B gene provided an important foundation for further studying the fatty deposition and lipornetabolism of FAM134 B gene in Luchuan pig.