人白细胞介素-10/人肝再生增强因子融合基因的构建及在肝细胞中的表达

Construction of human interleukin-10/augmenter of regeneration fused gene and its expression in hepa-tocyte

目的 利用人自细胞介索(IL)-10和肝再生增强因子(ALP)构建融合基因hIL-10／ALR和真核表达载体,验证其在肝细胞中的表达。方法 以富含疏水氨基酸(Gly-Ser)n的相应寡核苷酸接头序列为媒介,通过hIL-10和hALR克隆载体构建真核质粒表达载体pcDNA3hIL-10／ALR,纯化后转染L02肝细胞,并以逆转录-聚合酶链反应(RT-PCR)方法检测其表达。结果 成功构建hIL-10／ALR融合基因,其真核表达载体pcDNA3hIL-10／ALR可在L02肝细胞中有效表达。结论通过融合基因转导方法可能实现IL-10和ALR的各自功能,为肝纤维化、肝硬变的基因治疗探索有效手段。

Objective To construct the human interleukin-10/augmenter of regeneration fused gene and its eukaryotic expression plasmid vector and demonstrate its expression in L02 hepatocyte. Methods The human interleukin-10 gene and human augmenter of regeneration gene were ligated with a specially designed oligonucleotide, whose corresponding amino acid sequence was rich of hydrophobic amino acid residues (Gly-Ser)n. L02 hepatocytes were transfected with the fused gene plasmid vector, and its expression was detected by RT-PCR. Results The hIL-10/ALR fused gene was successfully constructed and could be effectively expressed in L02 hepatocytes. Conclusion The hIL-10 and hALR gene may realize to show their corresponsive function with the fused gene transduction, and set up an effective approach to exploration of gene therapy of liver fibrosis and cirrhosis.