摘要
目的 为研究软骨细胞基因转染的可行性创造条件 ,从而为基因修饰的软骨组织工程研究奠定基础。方法 应用基因重组技术和限制性内切酶酶切构建并鉴定 pcDNA3 .1 TGF β1真核表达载体 ,经Fugene6介导质粒转染 3T3细胞后应用逆转录 聚合酶链反应 (RT PCR)检测hTGF β1mRNA在真核细胞中的表达。结果 重组真核表达载体 pcDNA3 .1 TGF β1经限制性内切酶XhoⅠ和HindⅢ酶切 ,电泳后显示 1 .35kb的hTGF β1目的片段和 5 .40kb的 pcDNA3 .1载体片段 ,证明重组质粒连接正确 ;经RT PCR检测了hTGF β1在转录水平mRNA的表达情况 ,表明质粒转染 3T3细胞后hTGF β1mRNA的表达明显增强。 结论 重组真核表达载体构建正确 ,并能在真核细胞 3T3中表达hTGF β1mRNA 。
Objective To analyze the biologic function of transforming growth factor β1 (TGF β1) and to study the possibility of gene transfection to chondrocytes.Methods A eukaryotic expression vector for TGF β1 (pcDNA3.1 TGF β1) was constructed by use of recombinant DNA technique and was transfected into 3T3 cells by using Fugene6.The mRNA expression of TGF β1 was detected by the RT PCR method.Results The recombinant eukaryotic expression vector for TGF β1 was digested with XhoⅠand HindⅢ,and the electrophoresis of the digested products showed two fragments:1.35?kb fragment and 5.40?kb fragment.The hTGF β1 gene transfected 3T3 cells showed prominently elevated mRNA expression of hTGF β1.Conclusion The recombinant vector was constructed correctly and the mRNA of human TGF β1 could be prominently expressed in 3T3 cells,which establish the base for hTGF β1 genetically augmented tissue engineering of the cartilage.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2002年第3期270-271,共2页
Chinese Journal of Experimental Surgery
基金
国家重点基础研究发展规划"973"项目(G19990 54 30 0 )
