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绵羊生殖道中ghrelin基因的克隆及其表达
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吕东媛，曹贵方*，李淑凤，赵鹏伟
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(内蒙古农业大学 动物科学与医学学院,内蒙古 呼和浩特010018)
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摘要：
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从雌性绵羊输卵管、子宫体、子宫颈、阴道组织中提取总RNA,根据已获得的绵羊ghrelin基因cDNA序列设计特异性引物,采用RT-PCR方法扩增出了绵羊ghrelin基因；将扩增产物克隆于pMD19-T载体后进行测序。以β-肌动蛋白(β-actin)基因作为内参,采用半定量RT-PCR法扩增ghrelin基因，经琼脂糖凝胶电泳后,应用凝胶成像分析系统计算雌性绵羊不同生殖道组织中ghrelin基因的表达量。结果显示,从雌性绵羊生殖道各组织均扩增出了233bp的ghrelin基因片段；ghrelin基因在子宫体的表达量最高，输卵管内次之，子宫颈和阴道内最低。表明，ghrelin对雌性绵羊生殖系统的调节及生殖激素的分泌等具有重要作用。 |
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关键词：
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ghrelin；绵羊；β-肌动蛋白；组织表达 |
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中图分类号：S 814.1 文献标识码：
A 文章编号：1673-4696(2008)03-0244-05 |
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Cloning and expression of ghrelin gene in genital tracts of sheep |
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L Dong-yuan,CAO Gui-fang,LI Shu-feng,ZHAO Peng-wei |
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(College of Animal Science and Medicine,Nei Monggol Agricultural University,Hohhot 010018,China)
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Abstract: Total RNAs were extracted from fallopian tube,uterus,cervix of uterus and vagina of female sheep,respectively.Based on the published ghrelin cDNA sequence of sheep,one pair of specific primers was designed,and the ghrelin gene of sheep was amplified by RT-PCR and cloned into pMD19-T vector and sequenced.Then，the β-actin gene was used as internal reference,and the expression level of ghrelin gene in different tissues of female ovine reproductive system was detected by the semi-quantitative RT-PCR amplification of the ghrelin gene and TANNONG gel image acquisition and analysis system software after agarose gel electrophoresis.Electrophoretic analysis showed that the ghrelin gene of 233bp in length was expressed in different tissues of genital tracts,with highest expression in uterus body,followed by fallopian tube,and the oviduct cervix of uterus and vagina had the lowest expression.These results indicated that ghrelin could regulate genital system and genital hormone secretion of female sheep. |
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Key words:ghrelin;sheep;β-actin;expression in tissue |
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