重组人生长激素基因的转染和基因表达产物的定量分析  

Transplantation of the transfected cells with recombinant hGH gene VI:gene transfection and quantitative analysis of gene expression product

作　　者：胡志强[1,2] 戴钦舜[1,2] 于康震[1,2] 杨富明[1,2] 刘恩重[1,2] 陈忠斌[1,2] 戴亚美[1,2] 辛晓敏[1,2] 高自强[1,2] 

机构地区：[1]哈尔滨医科大学第一临床医学院神经外科 [2]哈尔滨兽医研究所国家生物技术重点实验室

出　　处：《中华神经外科杂志》1998年第2期75-77,共3页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金

摘　　要：目的：定量分析基因表达产物人生长激素（ｈＧＨ），并确定基因表达期限。方法：我们利用磷酸钙介导的方法将重组ｈＧＨ基因导入培养的人胚皮肤成纤维细胞内，之后用Ｎｏｒｔｈｅｒｎ杂交证实转染细胞能够表达ｈＧＨ基因。在此基础上又采用放免法对ｈＧＨ进行了定量测定。结果：从细胞转染的第１２天算起，ｈＧＨ水平逐渐提高，到２３天进入一稳定表达阶段，到９０天仍在进行稳定表达。此时每毫升培养液中的细胞数为１．３×１０６个，ｈＧＨ含量为７９．３２±３．２６６３ｎｇ。结论：转染细胞至少能持续表达ｈＧＨ９０天。Objective: To quantitative analyze the gene expression producthGH, and determine expression time limit. Methods: we use Northern blotting to prove the human fetal skin fibroblasts (hSF), which were transfected with recombinant hGH gene by calcium phosphate transfection system, could express the foreign hGH gene. On the basis of above researches, we adopted radioimmunoassay (RIA) for quantitative analyze of hGH. Results: hGH content gradually increased on the 12th day after transfection. The gene expression began to enter the steady expression stage on the 23rd day, and still expressed sustainedly hGH on the 90th day. During the stage, there were 13×106 hSF in a milliliter medium, hGH content was 7932±32663ng/ml. Conclusions: The transfected hSF can express hGH sustaintedly for at least 90 days.

关 键 词：HGH 基因转染 基因治疗 生长激素 侏儒症 

分 类 号：R584.210.5[医药卫生—内分泌]