中文摘要：

目的构建人转化生长因子-β1（TGFβ1）基因RNAi质粒载体。方法根据人TGFβ1 mRNA序列选择3个靶序列并设计合成相应3对寡核苷酸序列，同时合成1对阴性对照寡核苷酸序列；将以上4对寡核苷酸序列退火后连入pRNA-U6．2／Lenti质粒并分别命名为pRNA-U6．2／Lenti-1、pRNA-U6．2／Lenti-2、pRNA-U6．2／Lenti-3、pRNA-U6．2／Lenti-4。酶切和测序鉴定后，将以上重组质粒转染Hela细胞，运用RT-PCR和ELISA检测TGFβ1 mRNA和蛋白的表达水平。结果酶切和测序证实目的寡核苷酸片段已被准确克隆到pRNA-U6．2／Lenti质粒；与对照组相比，pRNA-U6．2／Lenti-1、pRNA-U6．2／Lenti-2转染Hela细胞后，TGFβ1 mRNA水平及蛋白水平的表达量均受到明显抑制；其中TGFβ1蛋白水平在pRNA-U6．2／Lenti-1组下降79．2％，在pRNA-U6．2／Lenti-2组下降53．7％，统计有显著性差异（P〈0．01）。结论成功构建了人TGFβ1基因RNAi质粒载体，对于TGFβ1高表达疾病的基因治疗奠定了基础。

英文摘要：

Objective To construct a plasmid vector of RNA interference （RNAi） of transforming growth factor-β1 gene （TGF-β1）, Methods First three target sequences were selected according to TGF-β1 mRNA sequence firstly, then three pairs of oligonucleotide sequences and one pair of negative control oligonucleotide sequence were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pRNA-U6.2/Lenti plasmid, which was then named pRNA-U6. 2/Lenti-1, pRNA-U6. 2/Lenti-2, pRNA-U6. 2/Lenti-3 and pRNA-U6. 2/Lenti-4, respectively. After being identified by restriction enzyme digestion and sequencing, the recombinant plasmids were transfected into Hela ceils. TGF-β1 expression in the transfected ceils was assayed by RT-PCR and ELISA. Results Enzyme digestion and DNA sequencing showed that the oligonucleotide fragments were correctly inserted into pRNA- U6.2/Lenti plasmid, and TGF-β1 expression in the transfected cells was knocked down significantly by pRNA-U6.2/ Lenti-1 and pRNA-U6.2/Lenti-2 at both the protein and mRNA level compared with those in the control group. TGF-β1 protein was reduced by a percentage of 79.2% and 53.7% in pRNA-U6.2/Lenti-1 and pRNA-U6.2/Lenti-2 group, respectively, which was significantly different from that in the control group （P〈0.01）. Conclusion The RNAi plasmid vector of TGF-β1 was constructed successfully, which laid foundation for gene therapy for diseases with high TGF-β1 expression.