猪11β-HSD1真核过表达载体的构建及其在Hepa1-6细胞中的表达

Abstract

Abstract: Glucocorticoids can be produced from inactive 11-keto forms through the enzyme 11β-hydroxysteroid dehydrogenase type 1. The cDNA of porcine 11β-HSD1 gene was amplified from the tissue of landrace pigs liver by RT-PCR and cloned into the eukaryotic expression vector pcDNA3.1. The recombinant was further identified by PCR amplification,enzyme digestion and DNA sequencing. The recombinant plasmid 11β-HSD1-pcDNA was transfected into the Hepa1-6 cell and its expression intensitye was detectable by RT-PCR. The results showed that the eukaryotic exprssion vector 11β-HSD1-pcDNA was successfully constructed which could provide a necessary basis for the research of transgenic pigs.

Key words: 11β-HSD1; eukaryotic expression; gene cloning; vector construction; transgenic

CLC Number:

XIA Ji-han, WANG Chao-qun, RUAN Jin-xue, YANG Shu-lin, LI Kui. Construction of 11β-HSD1 Eukaryotic Overexpression Vector and its Expression in Hepa1-6 Cell[J]. , 2012, 39(12): 1-5.

References

1 万小莉,伍伟.11β-羟基类固醇脱氢酶1与2型糖尿病[J].重庆医学,2007,19(36):2016~2018.
2 王继荣,许红霞,陈刚,等. 11β-羟基类固醇脱氢酶在人肠癌中的表达[J].现代肿瘤医学,2007,15(11):1619~1622.
3 邵喜英,陈占红,曹江,等. pcDNA3.1(+)-CYP19-GFP真核表达质粒构建[J].浙江大学学报(医学版),2011,40(2):189~194.
4 綦雯雯,钟历勇. 11β-羟基类固醇脱氢酶1型与认知功能[J].国际内分泌代谢杂志,2010,30(4):280~282.
5 程庆丰,李启富. 11β-羟类固醇脱氢酶1与代谢综合征关系研究进展[J].国外医学(内科学分册),2005,32(12):525~529.
6 Campino C,Quinteros H,Owen G I,et al. 11β-hydroxysteroid dehydrogenase type 2 polymorphisms and activity in a Chilean essential hypertensive and normotensive cohort[J]. Am J Hypertens, 2012,25(5):597~603.
7 Dujic T T,Bego T,Mlinar B,et al. Association between 11beta-hydroxysteroid dehydrogenase type 1 gene polymorphisms and metabolic syndrome in Bosnian population[J]. Biochem Med(Zagreb), 2012,22(1):76~85.
8 Masuzaki H,Paterson J,Shinayama H,et al. A transgenic model of visceral obesity and the metabolic syndrome[J].Science, 2001,294(5549):2166~2170.
9 Paterson J M,Morton N M,Fievet C,et al. Metabolic syndrome without obesity: Hepatic overexpression of 11β-hydroxysteroid dehydrogenase type 1 in transgenic mice[J]. Proceedings of the National Academy of Sciences of the United States of America,2004,101(18):7088~7093.
10 Pereira C D,Azevedo I,Monteiro R,et al. 11β-hydroxysteroid dehydrogenase type 1: Relevance of its modulation in the pathophysiology of obesity, the metabolic syndrome and type 2 diabetes mellitus[J]. Diabetes Obes Metab,2012,14(10):869~881.
11 Turban S,Liu X,Ramage L,et al. Optimal elevation of beta-cell 11beta-hydroxysteroid dehydrogenase type 1 is a compensatory mechanism that prevents high-fat diet-induced beta-cell failure[J]. Diabetes,2012,61(3):642~652.
12 Xia G,Liu L,Xue M,et al.Discovery of novel sulfonamides as potent and selective inhibitors against human and mouse 11beta-hydroxysteroid dehydrogenase type 1[J]. Mol Cell Endocrinol,2012,358(1): 46~52.

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Construction of 11β-HSD1 Eukaryotic Overexpression Vector and its Expression in Hepa1-6 Cell

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