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在人类的一些肿瘤中已发现存在Gsα基因突变。本研究室在白血病细胞系的研究中发现了Gsα基因的 3个缺失突变体以及野生型的Gsα(分别命名为GsαL 1,5 0 0bp ;GsαL 2 ,30 0bp ;GsαL 3,2 0 0bp和Gsα 4 ,12 0 0bp) ,并已在一些急性白血病患者中检测到。为进一步研究这些缺失体的结构、功能和生物学意义 ,作者将克隆了GsαL 1,GsαL 2和Gsα 4基因的质粒分别转化到大肠杆菌DH5α中 ,提取DNA ,采用特异引物进行PCR扩增 ,获得相应的目的基因 ,分别将其克隆到 pET2 2b(+ )表达载体上 ,再转化大肠杆菌进行表达。经培养 ,取菌体行SDS PAGE电泳分析。研究结果 :3个基因都有相应分子量的蛋白表达 ,且均以包涵体形式存在。这表明构建的这 3个基因确具完整的基因结构 ,及相应的蛋白表达。本研究为进一步研究它们的功能及其与白血病发生的关系打下了基础
In some human tumors have been found to have Gsα gene mutations. In this study, three deletion mutants of Gsα gene and wild-type Gsα (named GsαL 1,5 0 0 bp; GsαL 2, 30 0 bp; GsαL 3,2 0 0 bp and Gsα 4, 1200bp) and has been detected in some acute leukemia patients. To further investigate the structural, functional and biological significance of these deletions, we cloned the plasmids of GsαL 1, GsαL 2 and Gsα 4 into Escherichia coli DH5α. DNA was extracted and amplified by specific primers to obtain The corresponding target genes were cloned into pET2 2b (+) expression vector and then transformed into E. coli for expression. After culture, take the bacteria SDS PAGE electrophoresis analysis. The results showed that all three genes had the corresponding molecular weight protein expression, and all existed as inclusion bodies. This indicates that these three genes are indeed constructed with a complete genetic structure, and the corresponding protein expression. This study laid the foundation for further study of their function and their relationship to leukemia