摘要
为研究牦牛雌激素受体(ERα和ERβ)基因序列特征及其在不同组织中的表达差异.根据NCBI上已公布的普通牛ERα和ERβ基因序列,设计特异性引物,利用RT-PCR技术克隆麦洼牦牛ERα和ERβ基因序列,同时利用荧光定量PCR技术检测ERα和ERβ基因在牦牛不同组织的表达分布.结果表明,分别获得2 100 bp ERα(Gen Ban K登录号:KJ011123)和1 791 bp ERβ(Gen Ban K登录号:KJ011124)基因序列,其中ERα的ORF为1 658 bp,编码596个氨基酸,ERβ的ORF为1 584 bp,编码527个氨基酸.多重序列分析表明,牦牛ERα和ERβ与普通牛、原鸡、人、小鼠、大鼠、扬子鳄、蟾蜍及斑马鱼的氨基酸序列同源性分别为45.3%-99.5%和53.9%-99.1%.荧光定量PCR结果显示,ERα和ERβ基因在牦牛的各种组织中均有表达,且除睾丸外,ERα在输卵管、子宫及乳腺中的表达高于ERβ.此外,就ERα而言,ERα在牦牛乳腺中表达最高,子宫、输卵管及卵巢次之,心、肝、脾、肺、肾及睾丸相对表达较低.而牦牛ERβ基因在卵巢中表达最高,子宫和输卵管次之,心、肝、脾、肺、肾、睾丸及乳腺表达较低.为进一步了解牦牛ERα和ERβ基因的生物学功能及其分子繁殖机制提供了基础.
The objective of this study is to explore the molecular characteristics and expression pattern of ERα and ERβ gene in yak(Bos grunniens). A pair of special primes were designed according to released sequence of bovine ERα and ERβ in Gen Bank. Then reverse transcription-polymerse chain reation(RT-PCR) was used to clone ERα and ERβ genes of Mai Wa yak, and Real-time PCR was used to identify the m RNA expression of ERα and ERβ. The result showed that Mai Wa yak ERα gene c DNA was 2 100 bp, with an ORF of 1 658 bp encoding 596 amino acids. ERβ gene c DNA was 1 791 bp, with an ORF of 1 584 bp encoding 527 amino acids. The sequence multialign results showed that yak ERα and ERβ gene shared 45.3%-99.5% and 53.9%-99.1% of similar amino sequence, with that of Bos Taurus, gallus gallus, Homo sapiens, Mus musculus, Rayyus norvegicus, alligator, xenopus laevis, and zebrafish. Real-time PCR analysis revealed that the ERα and ERβ were expressed in a variety of tissues, but the expression of ERα was higher than ERβ in productive organs. Additionally, as far as ERα, the m RNA expression was the highest in mammary gland, followed by uterus, oviduct and ovary, and lowest in liver, kidney, lung, testis, spleen and heart. The ERβ m RNA level was highest in ovary, intermediary in uterus and oviduct, and lowest in heart, liver, spleen, lung, kidney, mammary gland and testis. Identification and tissue distribution of yak ERα and ERβ genes provided foundation for the further study on their biological roles in yak.
引文
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