肝癌相关基因DLK1转基因小鼠的构建与鉴定

目的 构建并鉴定肝脏特异性表达的DLKl转基因小鼠.方法 通过基因重组方法,将小鼠DLK1 cDNA片段置于小鼠白蛋白基因增强子和启动子序列下游,构建肝脏特异性表达的DLKl重组质粒,酶切重组质粒得到转基因片段,转基因在体外进行表达鉴定后,显微注射获得DLK1转基因首建小鼠,对首建小鼠进行传代,利用F1代小鼠进行DLK1转基因表达的鉴定.结果 逆转录(RT)-PCR和细胞免疫荧光显示,转基因DLK1片段可在小鼠肝癌细胞系Hep1-6中表达.RT-PCR和免疫组化结果显示,DLK1在F1代成年转基因小鼠肝脏中特异性表达.结论 成功构建了肝脏特异性表达的DLK1转基因小鼠.

Generation of transgenic mice with liver-specific expression of delta drosophila homolog-like 1

Objective To establish and identify transgenic mice model that specifically express delta drosophila homolog-like 1 (DLK1) in liver for the functional study of DLK1 on liver differentiation,hepatopathy,and hepatocellular carcinoma.Methods The coding sequence of DLK1 cDNA was inserted downstream of mouse albumin gene enhancer/promoter to construct a liver-specific DLK1 expression recombinant vector.The DNA fragment of transgene digested from the recombinant vector by Pine I was transfected to Hepl-6 cells to verify the expression of DLK1 in vitro.Then the transgene fragment was microinjeeted into fertilized eggs of C57×CBA F1 mice and 20 transgenic founder mice were generated.The F1 mice of DLK1 transgenie founder mice were used to identify the expression of the transgene in liver and other tissues.Results RT-PCR and cellular immunofluorescence showed DLK1 expression when the transgene fragment was transfeeted into Hep1-6.For transgenic mice,RT-PCR and immunohistochemistry showed that DLK1 was specifically expressed in adult F1 mice liver.Conclusion Successfully established the liver-specific DLK1 expression transgenic mice model.