| 正义、反义Id1真核表达载体的构建及其对胃癌细胞的转染 |
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| 引用本文： | 张捷,王川,郭雯珲,傅芳萌,洪天姿,郑美春.正义、反义Id1真核表达载体的构建及其对胃癌细胞的转染[J].福建医科大学学报,2009,43(4):288-291. |
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| 作者姓名： | 张捷 王川 郭雯珲 傅芳萌 洪天姿 郑美春 |
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| 作者单位： | 福建医科大学,附属协和医院肿瘤科,福州,350001 |
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| 摘 要： | 目的构建正义、反义Id1真核表达载体,并转染胃癌SGC7901细胞株,筛选稳定表达的细胞株。方法设计Id1基因两端的引物,RT-PCR扩增Id1全长序列。将目的基因插入pcDNA3.1＋/Hygro构建正义Id1真核表达载体,插入pcDNA3.1-/Hygro构建反义Id1真核表达载体。脂质体转染法转染胃癌SGC7901细胞系,潮霉素筛选稳定表达的细胞株。结果RT-PCR法获得长约530 bp的Id1全长序列,经酶切及测序鉴定正确后转染胃癌SGC7901细胞株。经过潮霉素筛选8周,获得抗性的细胞株。经Western-blot鉴定,转染正义Id1真核表达载体的细胞株Id1蛋白表达水平高于对照组,而转染反义Id1真核表达载体的细胞株Id1蛋白表达水平低于对照组。结论成功构建正义、反义Id1真核表达载体并转染胃癌SGC7901细胞株,为进一步研究打下基础。
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| 关 键 词： | 抑制因子 免疫 真核细胞 遗传载体 胃肿瘤 转染 |
Construction of the Sense and Antisense Id1 Eukaryotic Expression Vector and Transfection of the Gastric Caner Cell Line |
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| ZHANG Jie,WANG Chuan,GUO Wenhui,FU Fangmeng,HONG Tianzi,ZHENG Meichun.Construction of the Sense and Antisense Id1 Eukaryotic Expression Vector and Transfection of the Gastric Caner Cell Line[J].Journal of Fujian Medical University,2009,43(4):288-291. |
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| Authors: | ZHANG Jie WANG Chuan GUO Wenhui FU Fangmeng HONG Tianzi ZHENG Meichun |
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| Institution: | ZHANG Jie,WANG Chuan,GUO Wenhui,FU Fangmeng,HONG Tianzi,ZHENG MeichunDepartment of Oncology,The Affiliated Union Hospital,Fujian Medical University,Fuzhou 350001,China |
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| Abstract: | Objective To construct the sense and antisense eukaryotic expression vector of Id1 and transfect to the SGC7901 cell lines of gastric cancer and select the stable hygromycin resistant cell lines. Method Design primers targeting two extremes of Id1 gene,and amplify the full-length of Id1 gene by means of RT-PCR.Insert the amplified DNA into PCDNA3.1+/Hygro to generate the sense Id1 eukaryotic expression vector;and into PCDNA3.1-/Hygro to generate the antisense Id1 eukaryotic expression vector.The recombinant... |
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| Keywords: | suppressor factors immunologic eukaryotic cells genetic vectors stomach neoplasms transfection |
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