本文關(guān)鍵詞：人睪丸特異性基因TDRG1在恒河猴中同源序列的生物信息學(xué)分析及實(shí)驗(yàn)驗(yàn)證　出處：《中南大學(xué)》2010年碩士論文　論文類型：學(xué)位論文

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【摘要】： 目的：克隆人睪丸特異基因TDRG1在恒河猴中的同源基因序列,分析該基因在恒河猴多種組織中的表達(dá)譜,為深入研究TDRG1基因功能尋找動(dòng)物模型。 方法：以人TDRG1基因全長(zhǎng)序列作為查詢序列,利用BLASTN軟件檢索恒河猴基因組數(shù)據(jù)庫,查詢到的同源序列,運(yùn)用RT-PCR、Northern blot、Real-time、免疫組化等實(shí)驗(yàn)初探TDRG1同源基因在恒河猴睪丸組織中以及在恒河猴全身多組織中的表達(dá)。 結(jié)果：生物信息學(xué)分析表明恒河猴基因組中檢索到與TDRG1同源序列,同源性達(dá)94%。RT-PCR驗(yàn)證了恒河猴睪丸中的同源序列；Northern Blot分析了TDRG1在恒河猴組織中同源基因表達(dá)的表達(dá)譜,結(jié)果顯示只在恒河猴睪丸組織表達(dá)陽性,其余組織表達(dá)陰性；Real-time PCR結(jié)果提示TDRG1在恒河猴睪丸中同源基因有表達(dá),但在恒河猴腎臟、膀胱、肝臟等組織中無表達(dá)；免疫組化結(jié)果顯示抗人TDRG1單克隆抗體能特異結(jié)合恒河猴睪丸中的同源蛋白,而在其他組織中無TDRG1同源蛋白的表達(dá)。 結(jié)論：TDRG1基因在恒河猴睪丸中有同源基因表達(dá),為今后建立轉(zhuǎn)基因或基因敲除動(dòng)物模型提供了理論和實(shí)驗(yàn)依據(jù)。
[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.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R-332

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本文編號(hào)：1374196