为探究CYP17-I基因在绿鳍马面鲀繁殖周期中的作用及其与性激素水平变化的相关性,实验首先通过SmartTMRace技术克隆得到了绿鳍马面鲀CYP17-I c DNA全长序列1 580 bp,编码511个氨基酸。通过半定量RT-PCR技术检测了CYP17-I基因的时空表达,...为探究CYP17-I基因在绿鳍马面鲀繁殖周期中的作用及其与性激素水平变化的相关性,实验首先通过SmartTMRace技术克隆得到了绿鳍马面鲀CYP17-I c DNA全长序列1 580 bp,编码511个氨基酸。通过半定量RT-PCR技术检测了CYP17-I基因的时空表达,并采用放射免疫测定法检测雄鱼和雌鱼血清睾酮(T)和雌二醇(E2)的含量变化。结果发现,CYP17-I在卵巢、精巢和脑中表达量较高,其次为胃、心脏、脾脏、肾脏,在垂体和鳃中也检测到了微弱表达。在周期表达中发现,无论在卵巢还是精巢中,CYP17-I基因的表达水平与血清T含量密切相关,呈明显周期性变化,最高值均出现在5月份的卵细胞或精子成熟期,而与血清E2含量无显著的相关性。研究表明,CYP17-I在鱼类繁殖周期中对雄性激素睾酮的生成起关键的调控作用。展开更多
As a specific gene of fish,cytochrome P450c17-Ⅱ(CYP17-Ⅱ) gene plays a key role in the growth,development and reproduction level of fish.In this study,the single-stranded conformational polymorphism(SSCP) technique w...As a specific gene of fish,cytochrome P450c17-Ⅱ(CYP17-Ⅱ) gene plays a key role in the growth,development and reproduction level of fish.In this study,the single-stranded conformational polymorphism(SSCP) technique was used to characterize polymorphisms within the coding region of CYP17-Ⅱ gene in a population of 75 male Japanese flounder(Paralichthys olivaceus).Three single nucleotide polymorphisms(SNPs) were identified in CYP17-Ⅱ gene of Japanese flounder.They were c.G594A(p.G188R),c.G939A and c.G1502A(p.G490D).SNP1(c.G594A),located in exon 4 of CYP17-Ⅱ gene,was significantly associated with gonadosomatic index(GSI).Individuals with genotype GG of SNP1 had significantly lower GSI(P < 0.05) than those with geno-type AA or AG.SNP2(c.G939A) located at the CpG island of CYP17-Ⅱ gene.The mutation changed the methylation of exon 6.Indi-viduals with genotype AA of SNP2 had significantly lower serum testosterone(T) level and hepatosomatic index(HSI) compared to those with genotype GG.The results suggested that SNP2 could influence the reproductive endocrine of male Japanese flounder.How-ever,the SNP3(c.G1502A) located in exon 9 did not affect the four measured reproductive traits.This study showed that CYP17-Ⅱ gene could be a potentially useful candidate gene for the research of genetic breeding and physiological aspects of Japanese flounder.展开更多
文摘为探究CYP17-I基因在绿鳍马面鲀繁殖周期中的作用及其与性激素水平变化的相关性,实验首先通过SmartTMRace技术克隆得到了绿鳍马面鲀CYP17-I c DNA全长序列1 580 bp,编码511个氨基酸。通过半定量RT-PCR技术检测了CYP17-I基因的时空表达,并采用放射免疫测定法检测雄鱼和雌鱼血清睾酮(T)和雌二醇(E2)的含量变化。结果发现,CYP17-I在卵巢、精巢和脑中表达量较高,其次为胃、心脏、脾脏、肾脏,在垂体和鳃中也检测到了微弱表达。在周期表达中发现,无论在卵巢还是精巢中,CYP17-I基因的表达水平与血清T含量密切相关,呈明显周期性变化,最高值均出现在5月份的卵细胞或精子成熟期,而与血清E2含量无显著的相关性。研究表明,CYP17-I在鱼类繁殖周期中对雄性激素睾酮的生成起关键的调控作用。
基金supported by the Natural Science Foun-dation of Shandong Province,China(No.ZR2009DQ011)the New Teacher Special Fund of Doctor of the Ministry of Education of China(No.20090132120006)the Postdoctoral Innovative Projects of Shandong Province,China(No.200702039)
文摘As a specific gene of fish,cytochrome P450c17-Ⅱ(CYP17-Ⅱ) gene plays a key role in the growth,development and reproduction level of fish.In this study,the single-stranded conformational polymorphism(SSCP) technique was used to characterize polymorphisms within the coding region of CYP17-Ⅱ gene in a population of 75 male Japanese flounder(Paralichthys olivaceus).Three single nucleotide polymorphisms(SNPs) were identified in CYP17-Ⅱ gene of Japanese flounder.They were c.G594A(p.G188R),c.G939A and c.G1502A(p.G490D).SNP1(c.G594A),located in exon 4 of CYP17-Ⅱ gene,was significantly associated with gonadosomatic index(GSI).Individuals with genotype GG of SNP1 had significantly lower GSI(P < 0.05) than those with geno-type AA or AG.SNP2(c.G939A) located at the CpG island of CYP17-Ⅱ gene.The mutation changed the methylation of exon 6.Indi-viduals with genotype AA of SNP2 had significantly lower serum testosterone(T) level and hepatosomatic index(HSI) compared to those with genotype GG.The results suggested that SNP2 could influence the reproductive endocrine of male Japanese flounder.How-ever,the SNP3(c.G1502A) located in exon 9 did not affect the four measured reproductive traits.This study showed that CYP17-Ⅱ gene could be a potentially useful candidate gene for the research of genetic breeding and physiological aspects of Japanese flounder.
