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目的构建携带TGF-β1的重组慢病毒表达载体,为TGF-β1基因转染骨髓间充质细胞(BMSCs)的研究奠定基础。方法利用基因克隆技术将TGF-β1基因定向克隆至慢病毒载体,构建TGF-β1基因重组质粒,并进行PCR、酶切和测序鉴定,并通过脂质体LP2000的介导慢病毒293T细胞。结果采用基因克隆技术构建的TGF-β1重组慢病毒经PCR、酶切及测序鉴定完全正确,转染293T细胞能够正确表达。结论基因克隆技术能成功构建TGF-β1重组慢病毒载体,转染293T细胞后能够正确表达,为TGF-β1基因转染骨髓间充质细胞(BMSCs)诱导免疫移植耐受的研究奠定基础。
Objective To construct a recombinant lentiviral vector carrying TGF-β1 and lay the foundation for the study of TGF-β1 gene transfection into bone marrow mesenchymal cells (BMSCs). Methods TGF-β1 gene was cloned into lentiviral vector by gene cloning technique. The recombinant plasmids of TGF-β1 gene were constructed and identified by PCR, restriction enzyme digestion and sequencing. The lentivirus 293T cells were induced by lipofectamine 2000. Results The TGF-β1 recombinant lentivirus constructed by gene cloning technique was completely correct by PCR, restriction enzyme digestion and sequencing. 293T cells transfected with 293T cells were correctly expressed. Conclusion Gene cloning technique can successfully construct TGF-β1 recombinant lentiviral vector and can be correctly expressed after transfection into 293T cells, which lays the foundation for the study of TGF-β1 gene transfection into bone marrow mesenchymal cells (BMSCs) to induce immune graft tolerance.