CAJ | 학술논문

目的构建人胰岛素样生长因子1（IGF-1）的真核细胞表达质粒。方法用PCR方法从人的肝细胞cDNA文库中克隆出IGF-1cDNA，然后定向插入真核细胞表达载体pcDNA3中，并用脂质体方法转染COS7细胞。用ELISA法和人胚肺纤维母细胞以及NIH3T3纤维细胞增殖法分别测定转染细胞上清液中IGF-1的含量和生物活性。结果重组的真核细胞表达质粒pcDNA3-IGF-1所含的IGF-1cDNA序列和插入方向均正确，其转染的COS7细胞分泌较高浓度的IGF-1，并且具有明显促进纤维细胞增殖的能力。结论本实验所构建的重组真核细胞表达质粒pcDNA3-IGF-1能够高效表达有活性的IGF-1，对进一步研究IGF-1体内表达的生理和病理作用有一定意义。
목적 구건인이도소양생장인자1（IGF-1）적진핵세포표체질립。방법 용PCR방법종인적간세포cDNA문고중극륭출IGF-1cDNA，연후정향삽입진핵세포표체재체pcDNA3중，병용지질체방법전염COS7세포。용ELISA법화인배폐섬유모세포이급NIH3T3섬유세포증식법분별측정전염세포상청액중IGF-1적함량화생물활성。결과 중조적진핵세포표체질립pcDNA3-IGF-1소함적IGF-1cDNA서렬화삽입방향균정학，기전염적COS7세포분비교고농도적IGF-1，병차구유명현촉진섬유세포증식적능력。결론 본실험소구건적중조진핵세포표체질립pcDNA3-IGF-1능구고효표체유활성적IGF-1，대진일보연구IGF-1체내표체적생리화병리작용유일정의의。
Objective To construct eukaryotic expression plasmid for humaninsulin-like growth factor 1(IGF-1) . Methods IGF-1 cDNA was cloned by PCR from the human liver cDNA library， and inserted into eukaryotic expression vector pcDNA3 with gene recombinant technique， forming recombinant pcDNA3-IGF-1 plasmid. The latter was transfected into COS7 cells with liposome-mediated method．Concentration and activity of IGF-1 in the supernatant were detected by ELISA and fibro-blast proliferation assays respectively. Results IGF-1cDNA sequence and its insertion direction in recombinant pcDNA3-IGF-1 plasmid were are correct. The COS7 cells transfected with the pcDNA3-IGF-1 plasmid secreted high concentration of IGF-1，which markedly induced the proliferation of human fibroblast and NIH3T3 cells. Conclusion The recombinant pcDNA3-IGF-1 plasmid can express active IGF-1 with high efficiency， which facilitates the study of its physical and pathological roles in vivo.