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目的:构建大鼠CD36真核表达载体,并在293T细胞中表达。方法:应用RT-PCR技术,从大鼠肺泡巨噬细胞NR8383细胞提取的总RNA中,获得CD36基因编码序列片段,克隆至真核表达载体pEGFP-N1中,对重组质粒进行酶切和测序鉴定后,以脂质体介导法转染至293T细胞,通过荧光显微镜和Western blot检测其在293T细胞中的表达。结果:酶切和测序证明重组真核表达载体pEGFP-N1-CD36构建成功,荧光显微镜及Western blot确认目的基因序列在293T细胞中过表达。结论:成功构建大鼠CD36基因的重组真核表达载体pEGFP-N1-CD36,并在293T细胞中过表达。
Objective: To construct rat eukaryotic expression vector CD36 and express it in 293T cells. Methods: The coding sequence of CD36 gene was obtained from total RNA extracted from rat alveolar macrophages NR8383 by RT-PCR and cloned into the eukaryotic expression vector pEGFP-N1. The recombinant plasmids were digested and sequenced After transfected into 293T cells by liposome-mediated method, the expression of 293T cells was detected by fluorescence microscopy and Western blot. Results: The recombinant eukaryotic expression vector pEGFP-N1-CD36 was successfully constructed by restriction enzyme digestion and sequencing. The target gene sequence was over-expressed in 293T cells by fluorescence microscopy and Western blot. Conclusion: The recombinant eukaryotic expression vector pEGFP-N1-CD36 of rat CD36 gene was successfully constructed and overexpressed in 293T cells.