CAJ | 학술논문

目的构建microRNA-21(miR-21)慢病毒抑制载体,为研究miR-21在小鼠体内的功能及作用机制打下基础.方法利用miR-21前体并将其克隆入LV3 pGLV-H1-GFP质粒中,经酶切及测序鉴定,利用脂质体将鉴定的阳性重组LV3 pGLV-H1-GFP-miR-21抑制载体、PG-P1-VSVG和pCMV-dR 3个质粒转染到HEK-293 T 细胞,将所得病毒感染293 T细胞,检测病毒滴度,并将制备的病毒颗粒感染乳鼠心肌细胞和尾静脉注射小鼠,检测miR-21在小鼠体内的表达.结果酶切及测序结果证明成功构建了LV3 pGLV-H1-GFP-miR-21重组质粒,并成功包装成慢病毒,病毒滴度为2.4×109 TU/ml,重组病毒成功感染心肌细胞,同时在Balb/c小鼠心脏有表达.结论成功构建miR-21慢病毒抑制载体,获得高效表达miR-21的慢病毒颗粒,为miR-21的进一步研究奠定了基础.
목적구건microRNA-21(miR-21)만병독억제재체,위연구miR-21재소서체내적공능급작용궤제타하기출.방법이용miR-21전체병장기극륭입LV3 pGLV-H1-GFP질립중,경매절급측서감정,이용지질체장감정적양성중조LV3 pGLV-H1-GFP-miR-21억제재체、PG-P1-VSVG화pCMV-dR 3개질립전염도HEK-293 T 세포,장소득병독감염293 T세포,검측병독적도,병장제비적병독과립감염유서심기세포화미정맥주사소서,검측miR-21재소서체내적표체.결과매절급측서결과증명성공구건료LV3 pGLV-H1-GFP-miR-21중조질립,병성공포장성만병독,병독적도위2.4×109 TU/ml,중조병독성공감염심기세포,동시재Balb/c소서심장유표체.결론성공구건miR-21만병독억제재체,획득고효표체miR-21적만병독과립,위miR-21적진일보연구전정료기출.
Objective Construct microRNA-21 (miR-21) inhibition of lentivirus for studying its function and mechanism of miR-21 in mice. Methods The precursor of miR-21 was cloned into the LV3 pGLV-H1-GFP. The LV3 pGLV-H1-GFP-miR-21 recombinant plasmids, PG-P1-VSVG and pCMV-dR were transfected HEK-293 T cells.Virus titer was detected after infection of 293 T cells.Finally we evaluated miR-21 expression after that the virus infected mice cardiomyocytes and intravenously injected mice. Results The results showed that LV3 pGLV-H1-GFP-miR-21 recombinant plasmid was successfully constructed.The lentivirus titer was 2.4×109 TU/ml.The recombinant virus successfully infected myocardial cells and Balb/c mice heart. Conclusion We successfully constructed miR-21 lentivirus and obtained expression of miR-21 lentivirus particles. That laid foundations for the function of miR-21.