人类重组激活基因-2表达研究
发布时间：2018-11-29 10:01
【摘要】： 重组激活基因RAGs产物的正确表达,对于淋巴细胞成熟后的结构和功能有着重要的作用,是Ig和TCR多样性的主要原因,在免疫应答中发挥重要作用。RAGs基因的突变和异常表达可引起严重的免疫缺陷疾病。本文采用RT-PCR方法从人类RAGs表达细胞中扩增RAG-2编码蛋白质cDNA,插入到真核表达载体,构建RAG-2的真核表达载体,并在体外转染人类RAG-2非表达细胞系。表达载体DNA测序结果显示与GeneBank已知序列完全一致,RAG-2基因已被正确插入到表达载体中;在转染细胞中用RT-PCR法检测到RAG-2 mRNA的表达。RAG-2表达载体的成功构建,为进一步在人类淋巴细胞中表达出有功能的RAGs分子的研究奠定了基础、为探讨RAGs表达异常所致免疫缺陷病的基因治疗方法提供理论依据。
[Abstract]:The correct expression of recombinant activated gene RAGs products plays an important role in the structure and function of lymphocytes after maturation, and is the main reason for the diversity of Ig and TCR. The mutation and abnormal expression of RAGs gene can lead to serious immune deficiency disease. RAG-2 encoding protein cDNA, was amplified from human RAGs expression cells by RT-PCR method and inserted into eukaryotic expression vector. The eukaryotic expression vector of RAG-2 was constructed and transfected into human RAG-2 non-expression cell line in vitro. The results of DNA sequencing of the expression vector showed that the RAG-2 gene had been inserted into the expression vector correctly. The expression of RAG-2 mRNA was detected by RT-PCR method in transfected cells. The successful construction of RAG-2 expression vector laid a foundation for the further study of the expression of functional RAGs molecules in human lymphocytes. To provide theoretical basis for gene therapy of immunodeficiency due to abnormal expression of RAGs.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392
本文编号：2364692
[Abstract]:The correct expression of recombinant activated gene RAGs products plays an important role in the structure and function of lymphocytes after maturation, and is the main reason for the diversity of Ig and TCR. The mutation and abnormal expression of RAGs gene can lead to serious immune deficiency disease. RAG-2 encoding protein cDNA, was amplified from human RAGs expression cells by RT-PCR method and inserted into eukaryotic expression vector. The eukaryotic expression vector of RAG-2 was constructed and transfected into human RAG-2 non-expression cell line in vitro. The results of DNA sequencing of the expression vector showed that the RAG-2 gene had been inserted into the expression vector correctly. The expression of RAG-2 mRNA was detected by RT-PCR method in transfected cells. The successful construction of RAG-2 expression vector laid a foundation for the further study of the expression of functional RAGs molecules in human lymphocytes. To provide theoretical basis for gene therapy of immunodeficiency due to abnormal expression of RAGs.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392
【共引文献】
相关期刊论文 前2条
1 詹曦菁,秦晓勇;RAG蛋白在V(D)J重组中的作用[J];生物技术通讯;2001年03期
2 詹曦菁 ,秦晓勇;RAG蛋白在V(D)J重组中的作用[J];医学综述;2001年10期
相关硕士学位论文 前2条
1 刘旭;人类重组激活基因-1（RAG-1）的表达研究[D];吉林大学;2008年
2 张雪利;红笛鲷重组激活基因克隆及表达分析[D];广东海洋大学;2012年
，本文编号：2364692
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