Clone Of Alternative Polyadenylation Isoforms And Polymorphisms Of 3'UTR Of Bovine BBOX1 Gene

The bovine BBOX1 locus maps on the chromosome 15,where a quantitative trait loci(QTL)affecting meat quality and carcass traits have been previously described.In this study the isoforms of alternative polyadenylation of bovine BBOX1 gene were cloned by3'RACE,expression profile of bovine BBOX1 gene were analyzed by RT-PCR,polymorphisms of the 3'UTR of BBOX1 gene were detected by PCR-SSCP,RT-RFLP and sequencing.We also sought to determine the relationship between polymorphisms of the3'UTR of BBOX1 gene and meat quality and carcass traits of Chinese red cattle.The main results are as follows:(1)We found three isoforms of alternative polyadenylation(APA)of bovine BBOX1 gene in Chinese red cattle.The length of three different 3'UTRs APA transcript variants were 229 bp,664bp,and 678 bp,respectively.Accession numbers of three isoforms in NCBI are KX431577,KX431578,and KX431579,respectively.(2)Express of bovine BBOX1 gene was analyzed by Semi-Quantitative RT-PCR in various tissues of Chinese red cattle.Expression data demonstrate that the bovine BBOX1 gene expressed in selected tissues of interest,but its expression level is higher in rumen,liver,and kidney.(3)In Chinese red cattle population,polymorphisms of 3'UTR of bovine BBOX1 gene were detected by PCR-SSCP and sequencing.Four polymorphic loci were found in3'UTR of bovine BBOX1 gene,among which three were single nucleotide polymorphisms(SNPs),one was indel mutation.The SNP c.650T>C was located in distal poly A signal.It maked poly A signal in distal “AATAAA” changed into“AACAAA”.All polymorphisms loci of 3'UTR of bovine BBOX1 gene were genotype by PCR-RFLP in Chinese red population.The population genetics parameters,allele frequencies,genotype frequencies,polymorphism information content,effective number of allele,and heterozygosity,were calculated.(4)Association of polymorphisms of 3'UTR of bovine BBOX1 gene with meat quality and carcass traits in Chinese red cattle was analyzed using general liner model(GLM) programe.The BBOX1/Bbv I genotypes were significantly associated with muscle fiber diameter,shear force,net meat weight,and carcass weight(P<0.01).In BBOX1/Hinc II locus,a significant effect on muscle fiber diameter,shear force,net meat weight,and carcass weight also was revealed(P<0.01).The results will give the valuable information for improving meat quality and carcass traits in cattle.