摘要
目的 研究花生四稀酸在细胞中的释放及前列腺素的合成,构建与花生四稀酸释放相关的表达系统。即胞内型磷脂酶A2(cytosolic phospholipase cPLA2)cDNA-pRC/CMV。方法 从克隆载体pMT2用Sal1制备cPAL2cDNA;平末端连接cPAL2-pRC/CMV;转染鼠巨噬细胞及筛选正向阳性克隆;northern blot检测cPLA2 cDNA基因表达。结果
Objective To study the release of arachidonic acid and the biosynthesis of prostaglandins,the expression system of pRC/CMV inserting cytosolic phospholipase cDNA was constituted.Methods to prepare cPLA 2 cDNA from pMT 2 plasmid with Sal1;bluut end ligation of cPLA 2 cDNA-pRC/CMV;transfect the recombinant into maerophage-like cell and choose the sence subcloning;checc the expression of cPLA 2 cDNA by Northeen blot.Results The expression was showed on the autoradiography film.pRC/CMV is an effective vector for hcPLA 2 cDNA expression this work lays a foundation for further study on the expression of cPLA 2 and fransformation from unsaturation fatty acid to prostaglandins.
出处
《北华大学学报(自然科学版)》
CAS
2000年第3X期202-203,232,共3页
Journal of Beihua University(Natural Science)
基金
与美国弗吉尼亚大学合作项目