江苏大学学报（医学版）2011，Vol.21Issue(4)：285-288,4.
次黄嘌呤-鸟嘌呤磷酸核糖转移酶缺陷型EL/4细胞系的建立
Establishment of hypoxanthine guanine phosphoribosyltransferase deficient EL/4 cell line
摘要
Abstract
Objective: To construct a recombinant eukaryotic expression vector of hsa-mir-216 a and to express it in gastric cancer SGC7901/ DDP cells for detecting its effects. Methods: Primers were designed based on the pre-mir-216 a sequence in miRBase databasse; the pre-mir-216 a gene was amplified by PCR and cloned into pGenesil-1 vector. The gastric cancer SGC7901/DDP cells were transfected with pGenesil-1-hsa-mir-216 a vector and the expression of mature miR-216 a was examined in SGC7901/ DDP cells by semi-quantitative RT-PCR and real-time PCR. Results: The results of DNA sequencing showed that sequences of pGenesil-1 -hsa-mir-216 a vector were correct; semi- quantitative RT-PCR and real- time PCR indicated thatmature mir-216 a was over- expressed in SGC7901/ DDP cells after transfection of pGenesil-l -hsa-mir-216 a vector, real- time PCR indicated that the expression of mature mir-216 a in S/ D/mir-216 a cells was ( 63. 530 ±0. 159) folds that of S/ D/ HK cells( t= 1. 201 , P ＜0. 01) . Conclusion : The eukaryotic expression vector of hsa-mir-16 a was successfully constructed and effectively exrpressed in SGC7901/ DDP cells.关键词
乙基甲磺酸/8-氮杂鸟嘌呤/次黄嘌呤-鸟嘌呤磷酸核糖转移酶/突变/EL/4淋巴瘤细胞Key words
mir-216 a/ eukaryotic expression vector/ SGC7901/DDP cells/ transfection分类
医药卫生引用本文复制引用
龚文,禚娅,蒋砚秋,潘晓园,邵启祥..次黄嘌呤-鸟嘌呤磷酸核糖转移酶缺陷型EL/4细胞系的建立[J].江苏大学学报（医学版）,2011,21(4):285-288,4.基金项目
国家自然科学基金资助项目(30671984) （30671984）
江苏省自然科学基金资助项目(BK2008231) （BK2008231）
