目的克隆并分析籽鹅卵巢产蛋性能相关基因EST1的全长cDNA序列。方法利用实时荧光定量PCR技术对EST1基因在籽鹅产蛋前期与产蛋期卵巢中mRNA表达水平进行检测,并采用RACE(Rapid amplification of cDNAends,RACE)技术对该基因全长cDNA序列...目的克隆并分析籽鹅卵巢产蛋性能相关基因EST1的全长cDNA序列。方法利用实时荧光定量PCR技术对EST1基因在籽鹅产蛋前期与产蛋期卵巢中mRNA表达水平进行检测,并采用RACE(Rapid amplification of cDNAends,RACE)技术对该基因全长cDNA序列进行克隆,应用生物信息学预测方法对其编码的蛋白质进行分析。结果籽鹅产蛋期卵巢组织中EST1基因mRNA的表达水平显著高于产蛋前期(P<0.05)。经RACE技术获得EST1基因全长cDNA序列长1715bp,具有单一的完整开放阅读框(ORF,14～1318bp),推测编码蛋白含434个氨基酸残基,相对分子质量为107100,等电点为5.00。该蛋白为细胞质内蛋白,含3个跨膜螺旋,蛋白序列中含1个信号肽切割位点。结论经分子生物学软件进行蛋白质功能预测,初步确定EST1基因为籽鹅α-烯醇化酶蛋白基因,推测该基因可能参与籽鹅产蛋性能的分子调控。展开更多
The laying quail is a worldwide breed which exhibits high economic value. In our current study, the vas- oactive intestinal peptide receptor-1 （VIPR-1） was selected as the candidate gene for identifying traits of eg...The laying quail is a worldwide breed which exhibits high economic value. In our current study, the vas- oactive intestinal peptide receptor-1 （VIPR-1） was selected as the candidate gene for identifying traits of egg produc- tion. A single nucleotide polymorphism （SNP） detection was performed in 443 individual quails, including 196 quails from the H line, 202 quails from the L line, and 45 wild quails. The SNPs were genotyped using a polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）. Two mutations （G373T, A313G） were detected in all the tested quail populations. The associated analysis showed that the SNP genotypes of the VIPR-1 gene were sig- nificantly linked with the egg weight of G373T and A313G in 398 quails. The quails with the genotype GG always exhibited the largest egg weight for the two mutations in the H and L lines. Linkage disequilibrium （LD） analysis in- dicated that G373T and A313G loci showed the weakest LD. Seven main diplotypes from the four main reconstructed haplotypes were observed, indicating a significant association of diplotypes with egg weight. Quails with the hlh2 （GGGT） diplotype always exhibited the smallest egg weight and largest egg number at 20 weeks of age. The overall results suggest that the alterations in quails may be linked with potential major loci or genes affecting reproductive traits.展开更多
文摘目的克隆并分析籽鹅卵巢产蛋性能相关基因EST1的全长cDNA序列。方法利用实时荧光定量PCR技术对EST1基因在籽鹅产蛋前期与产蛋期卵巢中mRNA表达水平进行检测,并采用RACE(Rapid amplification of cDNAends,RACE)技术对该基因全长cDNA序列进行克隆,应用生物信息学预测方法对其编码的蛋白质进行分析。结果籽鹅产蛋期卵巢组织中EST1基因mRNA的表达水平显著高于产蛋前期(P<0.05)。经RACE技术获得EST1基因全长cDNA序列长1715bp,具有单一的完整开放阅读框(ORF,14～1318bp),推测编码蛋白含434个氨基酸残基,相对分子质量为107100,等电点为5.00。该蛋白为细胞质内蛋白,含3个跨膜螺旋,蛋白序列中含1个信号肽切割位点。结论经分子生物学软件进行蛋白质功能预测,初步确定EST1基因为籽鹅α-烯醇化酶蛋白基因,推测该基因可能参与籽鹅产蛋性能的分子调控。
基金supported by the Open Project of Hubei Key Laboratory of Animal Embryo and Molecular Breeding(No.2015ZD146),China
文摘The laying quail is a worldwide breed which exhibits high economic value. In our current study, the vas- oactive intestinal peptide receptor-1 （VIPR-1） was selected as the candidate gene for identifying traits of egg produc- tion. A single nucleotide polymorphism （SNP） detection was performed in 443 individual quails, including 196 quails from the H line, 202 quails from the L line, and 45 wild quails. The SNPs were genotyped using a polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）. Two mutations （G373T, A313G） were detected in all the tested quail populations. The associated analysis showed that the SNP genotypes of the VIPR-1 gene were sig- nificantly linked with the egg weight of G373T and A313G in 398 quails. The quails with the genotype GG always exhibited the largest egg weight for the two mutations in the H and L lines. Linkage disequilibrium （LD） analysis in- dicated that G373T and A313G loci showed the weakest LD. Seven main diplotypes from the four main reconstructed haplotypes were observed, indicating a significant association of diplotypes with egg weight. Quails with the hlh2 （GGGT） diplotype always exhibited the smallest egg weight and largest egg number at 20 weeks of age. The overall results suggest that the alterations in quails may be linked with potential major loci or genes affecting reproductive traits.
