中国医药
中國醫藥
중국의약
CHINA MEDICINE
2010年
6期
505-507
,共3页
文通%魏云峰%王梦洪%郑泽琪%彭景添%曾元临%曾俊义
文通%魏雲峰%王夢洪%鄭澤琪%彭景添%曾元臨%曾俊義
문통%위운봉%왕몽홍%정택기%팽경첨%증원림%증준의
骨髓间充质干细胞%miRNA-1%真核表达载体
骨髓間充質榦細胞%miRNA-1%真覈錶達載體
골수간충질간세포%miRNA-1%진핵표체재체
Bone marrow mesenchymal stem cells%miRNA-1%Eukaryotic expression vector
目的 构建大鼠miRNA-1真核表达载体,为研究miRNA-1功能提供实验基础.方法 应用技术体外合成miRNA-1前体对应的基因组片段,定向克隆到pSD11-U6/Neo/绿色荧光蛋白载体上.阳性克隆进行及DNA测序鉴定,将构建成功的重组质粒pSD11-U6/Neo/绿色荧光蛋白/miRNA-1借助脂质体转染大鼠骨髓间充质干细胞,荧光显微镜下观察绿色荧光蛋白表达.结果 PCR鉴定及DNA测序表明真核表达载体pSD11-U6/Neo/绿色荧光蛋白/miRNA-1构建成功.转染大鼠骨髓间充质干细胞后,荧光显微镜下观察到绿色荧光蛋白表达.结论 大鼠miRNA-1真核表达载体构建成功.
目的 構建大鼠miRNA-1真覈錶達載體,為研究miRNA-1功能提供實驗基礎.方法 應用技術體外閤成miRNA-1前體對應的基因組片段,定嚮剋隆到pSD11-U6/Neo/綠色熒光蛋白載體上.暘性剋隆進行及DNA測序鑒定,將構建成功的重組質粒pSD11-U6/Neo/綠色熒光蛋白/miRNA-1藉助脂質體轉染大鼠骨髓間充質榦細胞,熒光顯微鏡下觀察綠色熒光蛋白錶達.結果 PCR鑒定及DNA測序錶明真覈錶達載體pSD11-U6/Neo/綠色熒光蛋白/miRNA-1構建成功.轉染大鼠骨髓間充質榦細胞後,熒光顯微鏡下觀察到綠色熒光蛋白錶達.結論 大鼠miRNA-1真覈錶達載體構建成功.
목적 구건대서miRNA-1진핵표체재체,위연구miRNA-1공능제공실험기출.방법 응용기술체외합성miRNA-1전체대응적기인조편단,정향극륭도pSD11-U6/Neo/록색형광단백재체상.양성극륭진행급DNA측서감정,장구건성공적중조질립pSD11-U6/Neo/록색형광단백/miRNA-1차조지질체전염대서골수간충질간세포,형광현미경하관찰록색형광단백표체.결과 PCR감정급DNA측서표명진핵표체재체pSD11-U6/Neo/록색형광단백/miRNA-1구건성공.전염대서골수간충질간세포후,형광현미경하관찰도록색형광단백표체.결론 대서miRNA-1진핵표체재체구건성공.
Objective To construct an eukaryotic expression vector for mo-miRNA-1 to provide the experimental basis for miRNA-1 functions.Methods miRNA-1 genomic sequence was amplified by polymerase chain reaction(PCR) and cloned into pSD11-U6/Neo/GFP plasmid.Positive clones were identified with PCR and DNA sequencing, then mesenchymal-like stem cell were transfected pSD11-U6/Neo/GFP/miRNA-1 recombinant plasmid with Lipofectamine.Expression of green fluorescent protein was observed under a fluorescence microscope.Results PCR identification and DNA sequencing showed that the eukaryotic expression vector for miRNA-1 constructed successfully, green fluorescent protein expression was detected after transfection.Conclusion The eukaryotic expression vector for mo-miRNA-1 can be constructed successfully.
