CAJ | 학술논문

目的观察腺相关病毒和人γ-干扰素的重组体pWlG转染淋巴细胞及在细胞内表达情况.方法将pWlG以DNA-磷酸钙共沉淀法转染人淋巴细胞株H9,再分别用聚合酶链反应(PCR)、逆转录-PCR(RT-PCR)和酶联免疫吸附试验(ELISA)等方法在DNA、RNA和蛋白质水平研究该重组体导入细胞和在细胞内表达的情况.结果转染pWIG的H9细胞的DNA和RNA经PCR和RT-PCR检测均有504 bp的DNA条带;用ELISA可测出γ-干扰素的表达量为17.2 pg/3×106细胞.结论重组体pWlG用DNA-磷酸钙共沉淀法导入细胞后,转录HulFN-γmRNA,并翻译人γ-干扰素蛋白.为基因治疗的进一步研究打下基础.
목적 관찰선상관병독화인γ-간우소적중조체pWlG전염림파세포급재세포내표체정황.방법 장pWlG이DNA-린산개공침정법전염인림파세포주H9,재분별용취합매련반응(PCR)、역전록-PCR(RT-PCR)화매련면역흡부시험(ELISA)등방법재DNA、RNA화단백질수평연구해중조체도입세포화재세포내표체적정황.결과 전염pWIG적H9세포적DNA화RNA경PCR화RT-PCR검측균유504 bp적DNA조대;용ELISA가측출γ-간우소적표체량위17.2 pg/3×106세포.결론 중조체pWlG용DNA-린산개공침정법도입세포후,전록HulFN-γmRNA,병번역인γ-간우소단백.위기인치료적진일보연구타하기출.
Objective In order to study the transfection and expression of the recombinant pWIG of Adenc-associated virus and Human interferon-γ(HuIFN-γ)in human lymphocyte.Methods After trans-fecting pWIG into human lymphocyte via calcium phosphate,DNA,RNA and protein extracted from transfeeted cells were detected by polymerase chain reaction(PCR),reverse transcription PCR(RT-PCR)and ELISA.Results The 504 bp bands were detected from PCR and RT-PCR products of the DNA and RNA from the cells transfected,and HuIFN-γ protein were detected to be 17.2 pg/3 X 106 cells by ELISA.Conclusion The recombinant pWIG could produce HuIFN-γ mRNA after the transfection via cal-cium phosphate,and translate corresponding protein further.This lays ground for the further research of gene therapy.