QSA基因表达规律的研究及QSA抗原制备

发布时间：2018-11-06 10:59

【摘要】：长期以来精子被认为是转录静止的,其作用仅仅是将父本基因组DNA传递给卵子。但近年来研究证实成熟精子中存在大量复杂的mRNAs。通过基因芯片结合PCR的方法,在成熟精子中检测出大约3500-5000种的mRNAs。研究发现精子中的mRNAs在受精过程中会传递给卵子,并且持续稳定地存在,从而推测其可能在受精或胚胎早期发育过程中具有重要的作用。此外,另有研究表明精子中mRNAs在精子发生过程中选择性地被保留下来,它们可能会在成熟精子中翻译新生蛋白以替换降解的蛋白或者产生新的蛋白用于精子的获能。为了探究成熟精子中mRNA被选择性保留的机制,我们以从人精子SAGE库中克隆到的未知基因QSA为研究对象,从基因水平上检测QSA的表达规律及时空分布从而推测其可能的生物学作用,并制备QSA融合蛋白作为抗原。本研究采用RT-PCR和原位杂交法检测QSA在人和小鼠多种组织中的表达规律,并原核表达以及纯化QSA融合蛋白作为抗原。RT-PCR结果显示QSA在小鼠的睾丸、精子和受精卵中特异性表达,而在小鼠卵细胞、囊胚等组织中未检测到该基因的mRNA。原位杂交结果表明QSA在人的部分腺体组织(如睾丸、甲状旁腺等)中显著表达。根据基因QSA表达模式,推测其mRNA在获能精子中的选择性保留可能与某种激素的产生或分泌相关,并且可能参与精子发生或早期胚胎发育过程,从而为精子传递具有父系效应的mRNAs提供新的证据。
[Abstract]:Sperm has long been thought to be transcriptional stationary, only to transfer the paternal genomic DNA to the egg. But in recent years, studies have confirmed the existence of a large number of complex mRNAs. in mature spermatozoa About 3500-5000 species of mRNAs. were detected in mature spermatozoa by gene microarray combined with PCR. It has been found that mRNAs in spermatozoa is transmitted to the egg during fertilization and exists steadily, which suggests that it may play an important role in fertilization or early embryonic development. In addition, other studies have shown that mRNAs in spermatozoa is selectively preserved during spermatogenesis, and they may translate new proteins in mature spermatozoa to replace degraded proteins or produce new proteins for sperm capacitation. To explore the mechanism of selective retention of mRNA in mature spermatozoa, we studied QSA, an unknown gene cloned from human sperm SAGE library. The expression and temporal and spatial distribution of QSA were detected at the gene level in order to speculate its possible biological function. QSA fusion protein was prepared as antigen. In this study, RT-PCR and in situ hybridization were used to detect the expression of QSA in human and mouse tissues. Prokaryotic expression and purification of QSA fusion protein were used as antigens. RT-PCR results showed that QSA was present in the testis of mice. The mRNA. of the gene was not detected in mouse oocytes, blastocysts and other tissues. In situ hybridization showed that QSA was significantly expressed in some human glands (such as testis, parathyroid glands, etc.). Based on the QSA expression pattern, it is suggested that the selective retention of mRNA in capacitated sperm may be related to the production or secretion of certain hormones, and may be involved in spermatogenesis or early embryonic development. This provides new evidence for the transmission of patrilineal mRNAs by spermatozoa.
【学位授予单位】：上海交通大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392.1

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