本文選題：GIDRP88A + 選擇性剪接��； 參考：《四川大學(xué)》2005年碩士論文

【摘要】：生長抑制和分化相關(guān)蛋白88(growth inhibition and differentiation related protein 88,GIDRP88)基因是在研究抗癌藥物作用機理時,通過抑制消減雜交等技術(shù)獲得的,初步認為該基因與腫瘤分化、生長和細胞周期相關(guān)。實驗提示該基因存在不同的表達形式,可能是選擇性剪接體,但是還需要進一步證實。 本學(xué)位論文報道了一個與細胞生長和分化相關(guān)的新基因GIDRP88A的克隆和研究結(jié)果,GIDRP88A是未知功能基因GIDRP88的選擇性剪接形式。采用RT-PCR技術(shù)擴增GIDRP88,然后采用3'-RACE和5'-RACE方法,從乳腺癌細胞MDA-MB-231擴增到一個GIDRP88的新剪接型全長cDNA,其全長為2416bp,命名為GIDRP88A。生物信息學(xué)分析表明,GIDRP88A基因的選擇性剪接位點都位于閱讀框內(nèi),其全長cDNA比GIDRP88短約1kb,其中6號外顯子缺失和5號外顯子部分缺失,并在5號外顯子缺失部位多出一個堿基A,導(dǎo)致終止密碼子提前出現(xiàn);GIDRP88A基因所編碼的蛋白質(zhì)與GIDRP88相比在C-端少了566個氨基酸殘基,而GIDRP88基因所編碼的蛋白質(zhì)在C-端有兩個與功能相關(guān)的重復(fù)區(qū),因此,可能導(dǎo)致GIDRP88A基因的功能部分喪失,但是,仍然保留了與細胞生長和分化相關(guān)的結(jié)構(gòu)域。此外,通過半定量RT-PCR技術(shù),分析了GIDRP88A以及GIDRP88在正常和腫瘤組織中的表達水平,發(fā)現(xiàn)GIDRP88A在正常組織中高表達,在腫瘤組織中則呈低水平表達;而GIDRP88在正常和腫瘤組織中均呈高表達,提示二者之間在功能作用
[Abstract]:The growth inhibition and differentiation related protein 88(growth inhibition and differentiation related protein 88 GIDRP88) gene was obtained by suppression subtractive hybridization while studying the mechanism of anticancer drugs. It is suggested that the gene is related to tumor differentiation, growth and cell cycle. The results suggest that the gene is expressed in different forms, and may be a selective splice, but it needs to be further confirmed. This dissertation reports the cloning and study of a novel gene GIDRP88A associated with cell growth and differentiation. The results show that GIDRP88A is a selective splicing form of the unknown functional gene GIDRP88. GIDRP88 was amplified by RT-PCR, and then a novel spliced full-length cDNAof GIDRP88 was amplified from the breast cancer cell MDA-MB-231 by 3'-RACE and 5'-RACE, which was named GIDRP88A with a full length of 2416bp. Bioinformatics analysis showed that the selective splicing sites of GIDRP88A gene were all located in the reading frame, and its full-length cDNA was about 1 kb shorter than that of GIDRP88, including exon 6 deletion and exon 5 partial deletion. At the deletion site of exon 5, there was an additional base A, which led to the early appearance of GIDRP88A gene, which was 566 amino acid residues less than that of GIDRP88 in C- terminus of GIDRP88A gene. The protein encoded by the GIDRP88 gene has two functional repeat regions at the C-terminal, which may result in partial loss of the function of the GIDRP88A gene, but still retain the domain related to cell growth and differentiation. In addition, the expression levels of GIDRP88A and GIDRP88 in normal and tumor tissues were analyzed by semi-quantitative RT-PCR technique. It was found that the expression of GIDRP88A was high in normal tissues and low in tumor tissues. However, the high expression of GIDRP88 in both normal and tumor tissues suggests that there is a functional role between them.
【學(xué)位授予單位】：四川大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R341

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