SNPs Identification Of Aquaporin 7 And LHβ Gene And Correlation Analysis On Semen Quality In Bulls

In this study ,genetic polymorphism of LHβand AQP7gene's 5s ites in 45 Simmental and Charolais cattles was detected by SSCP(polymerase chain reaction- Single Strand Conformation Polymorphism).Polymorphism fragment of LHβand AQP7gene was cloned and sequenced so as to ascertain variant location of mutational sites and base type. The correlativity between these polymorphism sites and cattle's semen quality was analysed, whose objective is that providing foundation of molecular biology for seed selection and selection about Simmental and Charolais cattles.The polymorphism of the exon1,2,3,4,5,7,8 and intron6 of Aquaporin7 gene in Simmental and Charolais cattles was analyzed by PCR-SSCP. The results showed that two alleles and two genotypes ,were detected in the exon2,3,4 and intron6 of the Aquaporin7 gene. The polymorphism fragments were cloned and sequenced.The sequencing results indicated that there was one single nucleotide mutation: A→G(1429) in the exon2. And led to the coding of amino acids (Asn→Ser) had change.The sequencing results indicated that there was one single nucleotide mutation:G→C(6348) in the exon3. And led to the coding of amino acids (Arg→Thr) had change.The sequencing results indicated that there was one single nucleotide mutation: G→A(13746) in the exon4. And led to the coding of amino acids (Gly→Arg) had change.The sequencing results indicated that there was one single nucleotide mutation: A missing(14639) in the intron6.The Chi-square test for goodness of fit showed that the frequencies of the Aquaporin7 genotypes in the population of Simmental and Charolais cattles agreed with Hardy-Weinberg equilibrium.The genetic correlations were obtained between genotypes of the exon2,3,4 and intron6 of Aquaporin7 gene and Semen quality by the least squareanalysis:the different exon2 genotypes significantly affected the acrosome integrity of the rate of frozen semenor CC genotype on Aquaporin7was significantly higher than for CD(P<0.01). And the ratio of live sperm frozen semen for CC genotype on Aquaporin7 was higher than for CD.( P<0.05). The different exon3 genotypes significantly affected the ratio of live sperm frozen semen for EE genotype on Aquaporin7 was significantly higher than for EF(P<0.01). and the acrosome integrity of the rate of frozen semen and frozen semen vitality for EE genotype on Aquaporin7 was higher than for EF.(P<0.05).The different exon4 genotypes were not significantly affected the Bovine Semen quality.The different intron6 genotypes significantly affected the ratio of live sperm frozen semen for OO genotype on Aquaporin7was significantly higher than for OP (P<0.01). Other traits were not significantly affected.The polymorphism of the exon2 of LHβgene in Simmental and Charolais cattles was analyzed by PCR-SSCP. The results showed that the polymorphisms were not detected by PCR-SSCP in the 5′UTR ,exon1 and exon3 of theLHβgene. And two alleles, A, and B, three genotypes AA ,ABand BB were detected in the exon2 of theLHβgene. The polymorphism fragments were cloned and sequenced. The sequencing results indicated that there was one single nucleotide mutation:G→A in the exon2 of the LHβgene. The Chi-square test for goodness of fit showed that the frequencies of the LHβgenotypes in the population of Simmental and Charolais cattles agreed with Hardy-Weinberg equilibrium.The genetic correlations were obtained between genotypes of the exon2 of LHβgene and semen quality by the least square analysis.The results indicated that different LHβgenotypes significantly affected the abnormal ejaculation and semen volume.It implied that Aquaporin7 gene and LHβgene should be a major gene or a QTL linked gene which associated with Bovine Semen quality.