人淋巴毒素基因在中国仓鼠细胞中表达

遗传 ›› 1991, Vol. 18 ›› Issue (3): 208-213.

人淋巴毒素基因在中国仓鼠细胞中表达

刘军涛;李昌本;倪铁华;邹益民;赵寿元

复旦大学遗传学研究所，上海 200433

收稿日期:1900-01-01 修回日期:1900-01-01

Expression of Human Lymphotoxin Gene in GHO-DHFR Cells

Liu Juntao, Li Changben, Ni Yiehua, Zhou Yiming, Zhao Shouyuan

Liu Juntao, Li Changben, Ni Yiehua, Zhou Yiming, Zhao Shouyuan

Received:1900-01-01 Revised:1900-01-01

摘要/Abstract

摘要： 将人淋巴毒素(HuLT)基因插入哺乳动物细胞表达载体质粒p91023,经磷酸钙-DNA共沉淀法转入中国仓鼠二氢叶酸还原酶缺陷型(CHO-DHFR-)细胞，获得了DHFR+细胞克隆。RNA点渍杂交分析和MTT染料还原测定淋巴毒素的生物活性，均证明HuLT基因在DHFR+细胞里已被转录和转译，并被分泌到细胞外。其杀伤靶细胞——L929细胞的生物活性不低于每毫升培养液200单位。

关键词: 二氢叶酸还原酶, p91-HuLT重组质粒, 细胞毒活性, 人淋巴毒素基因, 人淋巴毒素基因, 细胞毒活性, 二氢叶酸还原酶, p91-HuLT重组质粒

Abstract: A recombinant plasmid p91-HuLT was constructed with a human lymphotoxin (HuLT) g ene 2.4kb EcoRI fragment and a mammalian cell expression vector plasmid p91023.T he HuL'T EcoRI DNA fragment covers entire coding sequence and some 3 1 non-trans lated region sequence of the gene.p91-HuLT DNA was used to transfect Chinese ham ster cell line CHO-DHFR- cells by using calcium phosphate-DNA coprecipitation me thod,and DHFR+ trasfectants were obtained RNA dot blot hybridization analysis sh owed that the HuLT mRNA was produced by p91-HuLT in transfectants.The results of MTT dye reduction assay indicated that the DHFR+ cell lines we obtained constit utively synthesize and secrete the HuLT with the cytotoxic activity of at least 200 units per mL of medium.

Key words: Recombinant plasmid p91-Hu LT, Human lymphotoxin (LuLT) gene, Cytotoxic activity, DHFR

刘军涛，李昌本，倪铁华，邹益民，赵寿元. 人淋巴毒素基因在中国仓鼠细胞中表达[J]. 遗传, 1991, 18(3): 208-213.

Liu Juntao, Li Changben, Ni Yiehua, Zhou Yiming, Zhao Shouyuan. Expression of Human Lymphotoxin Gene in GHO-DHFR Cells[J]. HEREDITAS, 1991, 18(3): 208-213.