鸭RIG-1基因的PCR扩增及其序列分析

维甲酸诱导基因1(RIG-1)是细胞质中侦测病原相关分子模式的识别受体。论文旨在扩增北京鸭RIG-1编码基因,并对其进行序列分析。通过PCR方法,从北京鸭脾脏中扩增得到RIG-1基因cDNA,并使用LaserGene分子生物学分析软件进行序列分析。结果发现北京鸭的RIG-1基因cDNA开放阅读框全长2 802bp,编码933个氨基酸。结构预测发现鸭RIG-1结构与哺乳动物类似,N端有两个串联CARD区,中间为DExD/H解旋酶区,C端为抑制区,各功能区起重要作用的氨基酸较为保守。同源性及进化树分析发现鸭RIG-1与鹅和斑马鱼的同源性最高分别为93.7%、78.4%,与哺乳动物同源性高于50%,与其他鱼类的同源性低于50%。成功扩增出北京鸭RIG-1基因cDNA编码序列,为鸭RIG-1的深入研究奠定了基础。

Amplification and Sequence Analysis of Duck RIG-1 Gene

Retinoic acid inducible gene 1(RIG-1) is a cytosolic pattern recognition receptor that senses pathogen-associated molecular patterns(PAMPs).This experiment was conducted to obtain the coding genes of duck RIG-1 to analysis its sequence and predict its domain.The full length cDNA of duck RIG-1 was amplified by PCR,and the sequence was analyzed by the bioinformatics software.The sequence analysis indicated that the open reading frame(ORF) of duck RIG-1 was 2 802 bp in length,encoded 933 amino acids.The domain prediction revealed the expected tandem N-terminal CARD domains,a helicase domain,a DeXD/H box helicase domain and a C-terminal repressor domain,consistent with the mammalian structure.The amino acids that play key roles in every domain are conserved.Phylogenetic analysis indicated that the homologies of amino acid sequences of the amplified full length cDNA were 93.7% and 78.4% to domestic goose and zebra fish,respectively.The coding gene of Beijing duck RIG-1 was amplified and the structure of putative protein was expected,and all these laid the foundation for the further study of duck RIG-1.