p53蛋白单、多克隆抗体的制备及免疫组化的研究
发布时间：2018-08-07 18:32
【摘要】：人类p53基因存在于17p13.1染色体上,编码分子量为53kDa的核磷酸化蛋白,p53基因是迄今发现的与人类肿瘤相关性最高的基因。P53基因突变及其表达产物在肿瘤发生时常伴随发生,且p53表达者预后较差,提示其与肿瘤的发生密切相关,检测其基因突变株及表达产物有助于肿瘤的诊断。p53基因突变后导致蛋白的组化性质发生改变,蛋白的稳定性大大增加,可通过分子生物学方法、免疫组化法、血清学等多种方法分析。而免疫组化方法是目前临床上对肿瘤,特别是疑难肿瘤分析检测所采用的重要方法,可以利用它分析p53基因突变诱导表达的蛋白在肿瘤组织中的情况。 本实验利用本实验组构建的PQE-p53表达质粒,在大肠杆菌M15表达系统中进行p53表达。建立p53蛋白在原核生物中的表达纯化方法,制备纯化的p53蛋白及相对抗体。目前虽然用于p53抗原检测的单、多抗很多,但大多是进口产品。所以基于以上情况我们尝试制备出可用于检测的单、多克隆抗体对肿瘤石蜡切片进行免疫分析,与进口产品其相比价格会大大降低,有助于肿瘤p53基因表达和突变的研究。
[Abstract]:Human p53 gene exists on the 17p13.1 chromosome, and the p53 gene encoding 53kDa is the most relevant gene. P53 gene mutation and its expression products often occur in tumorigenesis. The prognosis of the patients with p53 expression was poor, which suggested that p53 gene mutation and its expression products were closely related to the occurrence of tumor. The detection of p53 gene mutation would help to change the histochemical properties of the protein and increase the stability of the protein. It can be analyzed by molecular biology, immunohistochemistry, serology and so on. Immunohistochemical method is an important method for the analysis and detection of tumor, especially difficult tumor. It can be used to analyze the expression of p53 gene mutation induced protein in tumor tissue. In this experiment, p53 was expressed in E. coli M15 expression system by using the PQE-p53 expression plasmid constructed in our experimental group. The method of expression and purification of p53 protein in prokaryote was established to prepare purified p53 protein and relative antibody. Although currently used for the detection of p53 antigen, many polyclonal antibodies, but mostly imported products. Therefore, we try to prepare a single, polyclonal antibody that can be used for detection of tumor paraffin sections. Compared with the imported products, the price will be greatly reduced, which is helpful for the study of p53 gene expression and mutation.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392
本文编号：2170959
[Abstract]:Human p53 gene exists on the 17p13.1 chromosome, and the p53 gene encoding 53kDa is the most relevant gene. P53 gene mutation and its expression products often occur in tumorigenesis. The prognosis of the patients with p53 expression was poor, which suggested that p53 gene mutation and its expression products were closely related to the occurrence of tumor. The detection of p53 gene mutation would help to change the histochemical properties of the protein and increase the stability of the protein. It can be analyzed by molecular biology, immunohistochemistry, serology and so on. Immunohistochemical method is an important method for the analysis and detection of tumor, especially difficult tumor. It can be used to analyze the expression of p53 gene mutation induced protein in tumor tissue. In this experiment, p53 was expressed in E. coli M15 expression system by using the PQE-p53 expression plasmid constructed in our experimental group. The method of expression and purification of p53 protein in prokaryote was established to prepare purified p53 protein and relative antibody. Although currently used for the detection of p53 antigen, many polyclonal antibodies, but mostly imported products. Therefore, we try to prepare a single, polyclonal antibody that can be used for detection of tumor paraffin sections. Compared with the imported products, the price will be greatly reduced, which is helpful for the study of p53 gene expression and mutation.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392
【参考文献】
相关期刊论文 前2条
1 郭秀君,于昕;p53蛋白的转录调节功能及其相应的结构特征[J];上海免疫学杂志;1999年01期
2 江南,吴开力,黄强,潘苏华;一种简便的考马斯亮蓝G250蛋白质染色方法[J];生物化学与生物物理进展;2000年05期
，本文编号：2170959
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