生命科学研究
生命科學研究
생명과학연구
LIFE SCIENCE RESEARCH
2010年
2期
137-140
,共4页
欧阳华伟%罗成群%周建大%卜平元
歐暘華偉%囉成群%週建大%蔔平元
구양화위%라성군%주건대%복평원
高迁移率族蛋白1%启动子%荧光素酶%报告基因
高遷移率族蛋白1%啟動子%熒光素酶%報告基因
고천이솔족단백1%계동자%형광소매%보고기인
HMGB1%promoter%luciferase%reporter genes
利用PCR技术扩增小鼠高迁移率族蛋白1(HMGB1)基因启动子序列,构建小鼠HMGB1启动子荧光素酶报告基因pGL3-basic-HMGB1.经PCR、酶切及测序鉴定后.用脂质体法将pGL3-basic-HMGB1转入巨噬细胞264.7中.并应用萤光素酶测定系统检测其活性.检测结果显示pGL3-basic-HMGB1具有启动子活性.小鼠HMGB1启动子荧光素酶报告基因pGL3-basic-HMGB1的成功构建,为进一步研究HMGB1提供基本材料.
利用PCR技術擴增小鼠高遷移率族蛋白1(HMGB1)基因啟動子序列,構建小鼠HMGB1啟動子熒光素酶報告基因pGL3-basic-HMGB1.經PCR、酶切及測序鑒定後.用脂質體法將pGL3-basic-HMGB1轉入巨噬細胞264.7中.併應用螢光素酶測定繫統檢測其活性.檢測結果顯示pGL3-basic-HMGB1具有啟動子活性.小鼠HMGB1啟動子熒光素酶報告基因pGL3-basic-HMGB1的成功構建,為進一步研究HMGB1提供基本材料.
이용PCR기술확증소서고천이솔족단백1(HMGB1)기인계동자서렬,구건소서HMGB1계동자형광소매보고기인pGL3-basic-HMGB1.경PCR、매절급측서감정후.용지질체법장pGL3-basic-HMGB1전입거서세포264.7중.병응용형광소매측정계통검측기활성.검측결과현시pGL3-basic-HMGB1구유계동자활성.소서HMGB1계동자형광소매보고기인pGL3-basic-HMGB1적성공구건,위진일보연구HMGB1제공기본재료.
In order to construct the luciferase reporter gene containing mouse HMGB1 promoter pGL3-basic-HMGB1, the promoter of high mobility group box 1(HMGB1) gene was amplified from mouse genomic DNA by Polymerase Chain Reaction (PCR). The recombinant PGL3-basic-HMGB1 was transfected into 264.7 cells by lipofectamine after it was confirmed by PCR analysis, restriction enzyme digestion, and DNA sequencing, then the activity of luciferase was detected. The result demonstrated that the PGL3-basic-HMGB1 had the promoter activity. The luciferase reporter gene containing mouse HMGB1 promoter pGL3-basic-HMGB1 is constructed successfully, and it will become essential material for further study.
