期刊论文

中文

中国动脉硬化杂志

1007-3949

2007

15

7

484-486

10.3969/j.issn.1007-3949.2007.07.002

中国博士后基金（2005038472） 教育部重点科技基金项目（104158） 湖南省自然科学基金（06jj5051）

本校为第一机构

目的 构建稳定表达基质细胞衍生因子1α的ECV304细胞系,研究基质细胞衍生因子1α在单核细胞/内皮细胞粘附中的作用.方法 将大鼠基质细胞衍生因子1α基因聚合酶链反应产物及质粒载体pcDNA3.1用EcoRⅠ进行酶切,去磷酸化,置于连接反应体系中进行连接,将连接产物氯化钙法转化DH5α菌.氨苄青霉素筛选并扩增阳性菌落,经酶切鉴定插入方向正确后,再用G418筛选、逆转录聚合酶链反应检测基质细胞衍生因子1α浓度,建立稳定转染ECV304细胞系.在6孔板上种植转染基质细胞衍生因子1α基因的ECV304细胞,加THP-1细胞37℃孵育30 min,磷酸缓冲液轻洗3遍,去除未粘附细胞,倒置显微镜下计数上、下、左、右、中5个视野,...

Aim To study the effect of stromal cell derived factor-1 alpha （SDF-1α） on adherence of ECV304/THP-1. Methods Rat SDF-1α gene and pcDNA3.1 were both cut with EcoRⅠ, dephosphorylated, and connected in connecting buffer. Then, the connecting product was transformed to DH5α by calcium chloride, scanned by ampicillin, and cut by enzyme for the right inserting identification, and sub-selected by G418, SDF-1α expression of ECV304 was measured by reverse transcription polymerase chain reaction （RT-PCR） to get a stabilized SDF-1α expressional...