脂肪甘油三酯脂酶（ATGL）载体构建
onstruction of adipose triglyceride lipase expressing vector.
目的：研究旨在构建脂肪甘油三酯脂酶（adipose triglyceride lipase,ATGL）基因的表达载体。 方法：调取C57BL/6小鼠的ATGL基因的cDNA文库，并使用高保真Taq酶克隆出含有相应酶切位点的ATGL的编码区全序列，并通过基因测序验证序列准确性。得到的目的基因片段利用T4链接酶接入pBluescriptⅡ质粒中，并再一次通过基因测序确定重组质粒连接的准确性。建好的质粒继而转染HEK293细胞，检测细胞中ATGL的mRNA和蛋白的水平。结果：PCR产物双酶切和基因测序确定连接正确，细胞中ATGL的mRNA和蛋白表达量显著增高。结论：本研究成功构建了C57BL/6小鼠ATGL基因的表达载体，建好的载体可用于ATGL基因功能的后续研究。
Objecive:The purpose of this study is to construct the expression vector of adipose triglyceride lipase. Methods:Retrieved the cDNA library of the ATGL gene of C57BL / 6 mice and cloned the coding region containing the corresponding restriction sites of the whole sequence of ATGL by using high-fidelity Taq enzyme. Then verified the accuracy of the sequence by gene sequencing. Connect the target gene and pBluescriptⅡplasmid through T4 ligase. Determined by gene sequencing accuracy to make sure that the sequence was correct. Then transfected the HEK293 cell with recombinant plasmid and detected the express level of ATGL mRNA and protein. Results:The PCR products of enzyme digestion and gene sequence were coincide with expectation. ATGL mRNA and protein expression in the cells was significantly increased. Conclusion: The results showed that the ATGL recombinant expressing vector was constructed successfully. This vector can be used to study the following function of ATGL gene.
李颖、吕琳、孙长颢、栗晓霞、李松涛
基础医学生物科学研究方法、生物科学研究技术生物化学
GL57BL/6小鼠载体构建
adipose triglyceride lipase(ATGL)C57BL/6 miceconstruction of expressing vector
李颖,吕琳,孙长颢,栗晓霞,李松涛.脂肪甘油三酯脂酶（ATGL）载体构建[EB/OL].(2014-05-20)[2025-08-02].http://www.paper.edu.cn/releasepaper/content/201405-344.点此复制
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