猪FcγRⅡB多种剪接异构体的基因克隆与序列分析

摘要

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FcγRⅡB DNA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.%0bjsctive :To investigate 1he po^moiphian s of porche FcyR E B generated by alternative splicing .M etiiods :RT-PCR was perfomed using a pair of prmeis designed accoiding to 1he sequence of published poicine FcyR IIB (DQ 026064) .Nucfeotide and anino acid sequence alignmentwas analyzed win DNAMAN software package .The structure of 1he sequence was characterfeed with CBS onlhe software package .D etailed anafysis of 1he published gencm ic sequence ibr 1he regie hcliding 1he CD 32 (FcyR II B ) bcus,obtained fern Sus scroia chtnn osan e 4 genan ic contig (NW -001886241.1) ,was carried out using 1he G enan eScan intern /exon predictiDn online softoare package .R e- suits :The six poicine FcyR IIB transcript variante have been cbned ,and 1he transcrpi2 and transcript bo1h lacked one alanine residue at signal peptide regiDns can pared ti transcriptl ,3 ,5 and 6 .The poicine FcyR IIB transcript 5 and 6 w ere too new transcript variante w ith onfy one ]g structural dan ain h extracellular region .The transcrjpt6 wasaputative soluble Fc acceptir. Conclusion :FcyR IIB exists po^m oiphian s by different selective splicing fern of RNA .