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| Construction of recombinant adenoviral vector overexpressing betatrophin |
| JIANG Dan1， XUE Hao-ying1， WANG Su1， YANG Qi-chao1， LIU Yuan-xin1， YANG Ling1， WANG Dong1，YIN Wei1， ZHUANG Ruo1，QIAN Wei-yun2， ZHAO Shuang-xia3， ZHOU Li-bin4， YUAN Guo-yue1 |
| （1. Department of Endocrinology，the Affiliated Hospital of Jiangsu University，Zhenjiang Jiangsu 212001； 2. Department of Gerontology，the Affiliated 3th Hospital of Suzhou University，Changzhou Jiangsu 213001； 3. Department of Endocrinology，Shanghai Jiaotong University Affiliated 9th People′s Hospital，Shanghai 200011； 4. Department of Endocrinology，Affiliated Ruijin Hospital of Medical School，Shanghai Jiaotong University，Shanghai 200025， China) |
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Abstract [Abstract]Objective: To construct and identify the recombinant adenoviral vector expressing mouse betatrophin gene by using gene homologous recombination and to provide an experimental tool for further studying the function of betatrophin. Methods: From the sequence of GenBank,betatrophin gene including of BamHⅠ/ AgeⅠrestriction enzyme cutting sites was obtained by chemical synthesis and was inserted into the GV314 vector(CMVMCS3FLAGSV40EGFP) carrying enhanced green fluorescent protein(EGFP) to construct shuttle plasmid pGV314betatrophin. The plasmid GV314betatrophin was linearized with BamHⅠ/ AgeⅠand transfered into adenovirus genomics plasmid pDC315 by using gene homologous recombination in vitro to construct recombinant adenovirus vector Adbetatrophin.The DNA of recombinant adenovirus vector was finally linearized and transfered into HEK293T cells to package recombinant adenovirus particles.Recombinant adenovirus particles were repeated amplification in 293T cells to check and identify supernatant of adenovirus by PCR, western blotting and sequencing technologies. Results: By application of PCR, western blotting and DNA sequencing, the positive recombinant adenovirus containing betatrophin gene could be transferred into HEK293T cells and were packaged successfully. Conclusion: The recombinant adenovirus vector carrying mouse betatrophin gene was successfully constructed
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Received: 17 December 2015
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| \[1\]王嫕,秦彦文.促代谢因子（Betatrophin）：一种新的活性多肽\[J\]. 生理科学进展，2014,45(2):118-120.\[2\]赵术君，王明明，刘师伟. Betatrophin与糖脂代谢\[J\]. 中华内分泌代谢杂志， 2015, 31(3):288-290.\[3\]Ren G, Kim JY, Smas CM. Identification of RIFL, a novel adipocyteenriched insulin target gene with a role in lipid metabolism\[J\]. Am J Physiol Endocrinol Metab, 2012,303(3):E334-E351.\[4\]Zhang R. Lipasin, a novel nutritionallyregulated liverenriched factor that regulates serum triglyceride levels\[J\]. Biochem Biophys Res Commun, 2012,424(4):786-792.\[5\]Quagliarini F, Wang Y, Kozlitina J, et al. Atypical angiopoietinlike protein that regulates ANGPTL3\[J\]. Proc Natl Acad Sci USA, 2012,109(48):19751-19756.\[6\]Yi P, Park JS, Melton DA. Betatrophin: a hormone that controls pancreatic β cell proliferation\[J\]. Cell, 2013,153(4):747-758.\[7\]袁国跃，胡浩，孙文君，等. 糖尿病与Betatrophin\[J\]. 中华内分泌代谢杂志，2014, 30(12):1041-1043.\[8\]Hu H, Sun W, Yu S, et al. Increased circulating levels of betatrophin in newly diagnosed type 2 diabetic patients\[J\]. Diabetes Care, 2014,37(10):2718-2722.\[9\]Zhang R, AbouSamra AB. Emerging roles of Lipasin as a critical lipid regulator\[J\]. Biochem Biophys Res Commun, 2013,432(3):401-405.\[10\]Gusarova V, Alexa CA, Na E, et al. ANGPTL8/Betatrophin does not control pancreatic β cell expansion\[J\]. Cell, 2014,159(3):691-696.\[11\]Yi P，Park JS，Melton DA. Perspectives on the activities of ANGPTL8/betatrophin\[J\]. Cell, 2014,159(3):467-468.\[12\]高志涛, 宋海岩, 朱茉莉. 人IL16基因重组腺病毒载体的构建与鉴定\[J\].细胞与分子免疫学杂志, 2010, 26(8):823-825. |
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2016, Vol. 26 
