人睾丸特异性基因TDRG1在恒河猴中同源序列的生物信息学分析及实验验证

发布时间：2018-01-03 14:05

本文关键词：人睾丸特异性基因TDRG1在恒河猴中同源序列的生物信息学分析及实验验证　出处：《中南大学》2010年硕士论文　论文类型：学位论文

更多相关文章： 睾丸基因 TDRG1 同源序列 恒河猴

【摘要】： 目的：克隆人睾丸特异基因TDRG1在恒河猴中的同源基因序列,分析该基因在恒河猴多种组织中的表达谱,为深入研究TDRG1基因功能寻找动物模型。方法：以人TDRG1基因全长序列作为查询序列,利用BLASTN软件检索恒河猴基因组数据库,查询到的同源序列,运用RT-PCR、Northern blot、Real-time、免疫组化等实验初探TDRG1同源基因在恒河猴睾丸组织中以及在恒河猴全身多组织中的表达。结果：生物信息学分析表明恒河猴基因组中检索到与TDRG1同源序列,同源性达94%。RT-PCR验证了恒河猴睾丸中的同源序列；Northern Blot分析了TDRG1在恒河猴组织中同源基因表达的表达谱,结果显示只在恒河猴睾丸组织表达阳性,其余组织表达阴性；Real-time PCR结果提示TDRG1在恒河猴睾丸中同源基因有表达,但在恒河猴肾脏、膀胱、肝脏等组织中无表达；免疫组化结果显示抗人TDRG1单克隆抗体能特异结合恒河猴睾丸中的同源蛋白,而在其他组织中无TDRG1同源蛋白的表达。结论：TDRG1基因在恒河猴睾丸中有同源基因表达,为今后建立转基因或基因敲除动物模型提供了理论和实验依据。
[Abstract]:Objective: to clone the homologous gene sequence of human testis specific gene TDRG1 in rhesus monkey and analyze its expression profile in various tissues of rhesus monkey. To search for animal models for further study on the function of TDRG1 gene. Methods: the full-length sequence of human TDRG1 gene was used as the query sequence, and the rhesus monkey genomic database was searched by BLASTN software. The homologous sequence was obtained and RT-PCR was used. Expression of TDRG1 homologous genes in testicular tissues of rhesus monkeys and in multiple tissues of rhesus monkeys. Results: bioinformatics analysis showed that the homologous sequence of TDRG1 was found in rhesus monkey genome, and the homology was 94%. RT-PCR confirmed the homologous sequence in testis of rhesus monkey. Northern Blot analyzed the expression profiles of homologous genes of TDRG1 in rhesus monkey tissues. The results showed that only positive expression of TDRG1 was found in testis of rhesus monkeys, but negative in other tissues. The results of Real-time PCR indicated that TDRG1 was expressed in the testis of rhesus monkeys, but not in the kidney, bladder and liver of rhesus monkeys. Immunohistochemical results showed that monoclonal antibodies against human TDRG1 could specifically bind homologous proteins in the testis of rhesus monkeys, but there was no expression of TDRG1 homologous proteins in other tissues. Conclusion there is homologous gene expression in the testis of rhesus monkey, which provides a theoretical and experimental basis for the establishment of transgenic or knockout animal models.
【学位授予单位】：中南大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R-332

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