慢病毒介导SOX9基因转染骨髓间充质细胞的实验研究骨科学专业毕业论文.docx

山西 山西医科大学硕士学位论文 II II Experimental study of Lentiviral vector of SOX9 gene transducing bone mesenehymal stem cells Abstract Objucive To construct one lentiviral vector containing mouse SRY-related high mobility group-box gene 9 (Sox9) and to transfect murine bone mesenehymal stem cells(mBMSCs) in vitro and observe the expression of target gene. Methods RNA from the vectors containing mouxe Sox9 gene were extracted and Sox9 genes were amplified by reverse transcription-Polymerase Chain Reaction (RT-PCR).The Sox9 genes were connected into lentiviral vectors pGC-FU.Then pGC-FU-Sox9 transduced into 293T cells to produce recombinant lentivirus called as Lenti-SOX9-EGFP. mBMSCs were transfected.The expression of target gene was detected by immunofluorescence,RT-PCR and Western blot. Results Lenti-SOX9-EGFP was recombined successfully and transduced efficiently into mBMSCs.The expression of Sox9 gene was confirmed by RT-PCR and Western blot. Conclusion Lentiviral vector of mouse SOX9 gene can transfected successfully into mBMSCs。Meanwhile，Sox9 gene may be expressed in Sox9 Transduced mBMSCs.This will provide target cells for the following study about Sox9 gene respairing cartilage injury. Keywords SOX9；Lentivirus；bone mesenehymal stem cells III III 英文缩略词 Abbreviations 英文缩写 英文全称 中文名称 bp base pair 碱基对 DMEM Dulbecco’s Modified Eagle’s Medium 培养基 OA osteoarthritis 骨关节炎 PCR Polymerase chain reaction 聚合酶链反应 cDNA Complementary deoxy nucleic acid 互补 DNA DMSO Dimethyl sulfoxide