牛BBOX1基因3'UTR变异体的克隆及其多态性分析

Cloning of 3'UTR Isoforms and Polymorphisms in Bovine BBOX1 Gene

为鉴定牛BBOX1基因选择性多聚腺苷酸化的不同变异体及其3'UTR的多态性,采用3'RACE的方法检测出BBOX1基因3个不同3'UTR全长的APA变异体,短APA变异体3'UTR长度为229 bp,长APA变异体3'UTR长度分别为664 bp和678 bp。利用SSCP与测序相结合的方法,在BBOX1基因的3'UTR中检测到10个多态性,其中7个为SNP,分别为c.12T>C、c.100A>G、c.241T>C、c.480T>C、c.557T>C、c.606T>C和c.650T>C;3个为插入或缺失突变,分别为c.28_29ins C、c.434_435del GT和c.512_513ins TGC。SNP c.650T>C定位在Poly(A)加尾信号PAS3中,使加尾信号序列AAUAAA突变成AACAAA,导致3'UTR长度为678 bp的变异体出现。

The objective of this study was to identify isoforms of 3' UTR and polymorphisms in 3' UTR regions of bovine BBOX1 gene.Three different alternative polyadenylation(APA) isoforms of 3' UTR in the BBOX1 gene were identified by 3' RACE.Their lengths were 229,664,678 bp,respectively.Ten polymorphisms loci were identified in 3' UTR of BBOX1 gene by single strand conformation polymorphism(SSCP) and sequencing.There were seven SNPs,including c.12 T>C,c.100 A>G,c.241 T>C,c.480 T>C,c.557 T>C,c.606 T>C,and c.650 T>C.Three were deletion/insertion mutations,including c.28_29ins C,c.434_435del GT,and c.512_513ins TGC.SNP c.650 T>C was located in polyadenylation signals sequence PAS3.Moreover,the mutation changed AAUAAA into AACAAA,resulting in presence of 678 bp isform of 3'UTR in bovine BBOX1 gene.