人CD40 ligand基因真核表达质粒的构建及对人肝癌细胞HepG2的促凋亡作用
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徐 智, 沈苏南, 钱晓萍, 禹立霞, 刘宝瑞
肝癌; CD40L; 质粒; 真核表达载体,人CD40ligand基因真核表达质粒的构建及对人肝癌细胞HepG2的促凋亡作用,徐智,沈苏南,钱晓萍,禹立霞,刘宝瑞
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徐智, 沈苏南, 钱晓萍, 禹立霞, 刘宝瑞, 南京大学医学院附属鼓楼医院肿瘤中心 江苏省南京市 210008
徐智, 女, 1980-06-19生, 江苏省南京市人, 汉族, 2003年南京大学医学院肿瘤学硕士生.
国家自然科学基金资助项目, No. 30471701
通讯作者: 刘宝瑞, 210008, 江苏省南京市中山路321号, 南京大学医学院附属鼓楼医院肿瘤中心. baoruiliu@medmail.com.cn
电话: 025-83106666-76382
收稿日期: 2005-08-29 接受日期: 2005-09-06
Construction and expression of a eukaryotic expression plasmid containing human CD40 ligand in human hepatocellular carcinoma cell line HepG2 and its effect on apoptosis
Zhi Xu, Su-Nan Shen, Xiao-Ping Qian, Li-Xia Yu, Bao-Rui Liu
Zhi Xu, Su-Nan Shen, Xiao-Ping Qian, Li-Xia Yu, Bao-Rui Liu, Cancer Center of Drum Tower Hospital, Medical School of Nanjing University, Nanjing 210008, Jiangsu Province, China
Supported by National Natural Science Foundation of China, No. 30471701
Correspondence to: Professor Bao-Rui Liu, Cancer Center of Drum Tower Hospital, Medical School of Nanjing University, 321 Zhongshan Road, Nanjing 210008, Jiangsu Province,China. baoruiliu@medmail.com.cn
Received: 2005-08-29 Accepted: 2005-09-06
Abstract
AIM: To construct and express a eukaryotic expression plasmid containing human CD40 ligand (CD40L) in human hepatocellular carcinoma cell line HepG2 for the biological function study of CD40L on HepG2 cells.
METHODS: Human CD40L cDNA was synthesized by reverse transcription-polymerase chain reaction (RT-PCR) with the specific primers from the RNA of human peripheral blood monocyte (PBMC) and directly ligated into the eukaryotic expression vector pcDNATM3.1/myc-His(-)A through digestion with specific restriction endonuclease. The recombined plasmid was transformed into the E. coli DH5a to amplify CD40L gene. Then HepG2 cells were divided into 4 groups. The cells in group A were transfected with the recombined plasmid, and group B with the blank plasmid (not containing CD40L cDNA). The cells in group C were just normally cultured, and the ones in group D weren’t transfected but added G418 as the control for transfection ......
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