人GIDRP88选择性剪接体的鉴定和功能研究

发布时间：2018-05-29 03:16

本文选题：GIDRP88A + 选择性剪接　；参考：《四川大学》2005年硕士论文

【摘要】：生长抑制和分化相关蛋白88(growth inhibition and differentiation related protein 88,GIDRP88)基因是在研究抗癌药物作用机理时,通过抑制消减杂交等技术获得的,初步认为该基因与肿瘤分化、生长和细胞周期相关。实验提示该基因存在不同的表达形式,可能是选择性剪接体,但是还需要进一步证实。本学位论文报道了一个与细胞生长和分化相关的新基因GIDRP88A的克隆和研究结果,GIDRP88A是未知功能基因GIDRP88的选择性剪接形式。采用RT-PCR技术扩增GIDRP88,然后采用3'-RACE和5'-RACE方法,从乳腺癌细胞MDA-MB-231扩增到一个GIDRP88的新剪接型全长cDNA,其全长为2416bp,命名为GIDRP88A。生物信息学分析表明,GIDRP88A基因的选择性剪接位点都位于阅读框内,其全长cDNA比GIDRP88短约1kb,其中6号外显子缺失和5号外显子部分缺失,并在5号外显子缺失部位多出一个碱基A,导致终止密码子提前出现;GIDRP88A基因所编码的蛋白质与GIDRP88相比在C-端少了566个氨基酸残基,而GIDRP88基因所编码的蛋白质在C-端有两个与功能相关的重复区,因此,可能导致GIDRP88A基因的功能部分丧失,但是,仍然保留了与细胞生长和分化相关的结构域。此外,通过半定量RT-PCR技术,分析了GIDRP88A以及GIDRP88在正常和肿瘤组织中的表达水平,发现GIDRP88A在正常组织中高表达,在肿瘤组织中则呈低水平表达;而GIDRP88在正常和肿瘤组织中均呈高表达,提示二者之间在功能作用
[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.
【学位授予单位】：四川大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R341

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