猪FcγR ⅡB多种剪接异构体的基因克隆与序列分析

Cloning and analysis of multiple porcine FcγR ⅡB transcript variants

目的:研究猪FcγRⅡB受体基因选择性剪接的多样性。方法:根据GenBank中猪FcγRⅡB(swFcγRⅡB)cDNA基因序列(DQ026064),应用Primer5.0软件设计合成了一对特异性引物,应用RT-PCR技术,从不同个体的猪肺泡巨噬细胞中扩增FcγRⅡB基因并进行测序分析,在GenBank中查找猪FcγRⅡBDNA序列(NW-001886241.1),并用相关软件分析其外显子组成。结果:共克隆出6个转录体,其中转录体2,4在信号肽部分都有一个丙氨酸的缺失。转录体5,6是两个新型的FcγRⅡB转录体,转录体5胞外区只有一个Ig结构域,转录体6是一个只含有一个Ig结构域的推导性可溶性Fc受体。结论:猪FcγRⅡB的表达因RNA选择性剪接方式不同而呈现出多样性。

Objective：To investigate the polymorphisms of porcine FcγR ⅡB generated by alternative splicing.Methods：RT-PCR was performed using a pair of primers designed according to the sequence of published porcine FcγR ⅡB（DQ026064）.Nucleotide and amino acid sequence alignment was analyzed with DNAMAN software package.The structure of the sequence was characterized with CBS online software package.Detailed analysis of the published genomic sequence for the region including the CD32（FcγR ⅡB） locus,obtained from Sus scrofa chromosome 4 genomic contig（NW-001886241.1）,was carried out using the GenomeScan intron/exon prediction online software package.Results：The six porcine FcγR ⅡB transcript variants have been cloned,and the transcript2 and transcript4 both lacked one alanine residue at signal peptide regions compared to transcript1,3,5 and 6.The porcine FcγR ⅡB transcript 5 and 6 were two new transcript variants with only one Ig structural domain in extracellular region.The transcript 6 was a putative soluble Fc acceptor.Conclusion：FcγR ⅡB exists polymorphisms by different selective splicing form of RNA.