猪胰岛素启动子序列分析及调控基因在胰腺组织特异性表达验证

doi:10.11843/j.issn.0366-6964.2013.10.002

Abstract

Abstract:

The analysis of different Chinese local pig breeds insulin promoter sequence provided the basis for studying miniature pig β cell function and preparing diabetes model. In this study, porcine insulin promoters（PIP）from three pig breeds(Tongcheng pigs, Guangxi Bama Miniature pigs and Wuzhishan Miniature pigs) had been amplified by PCR. By the comparative analysis, the sequences of local pig breeds characteristics were got. To verify the validities and specificities of the gene expression regulatory of the sequence, pancreatic tumor cells and transgenic mice were used. There were no differences among the three local pig breeds in 1.5 kb insulin promoter sequences. Compared with the sequence from foreign pig breeds, there were only three nucleotide differences. Dual luciferase reporter gene detection system proved that the promoter could efficiently start downstream gene expression; In transgenic mice with CHOP and UCP2, two target genes expressions in pancreatic tissue were significantly higher than that in the liver, back muscles and kidneys, and pancreas tissue protein abundance significantly increased. The result of this study shows that the insulin promoter of Chinese local pig breeds can regulate target gene expression in the pancreatic tissue, which lays the foundation for producing transgenic pig model of regulating β cell function.

CLC Number:

S828
S813.3

HU Bing-jun, WANG Chao-qun, RUAN Jin-xue, XIN Lei-lei, MOU Yu-lian,YANG Shu-lin, LI Kui. Analysis of the Porcine Insulin Promoter and Verification of the Regulatory Genes Specific Expression in Pancreatic Tissue[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(10): 1516-1521.

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References

［1］YANG W, LU J, WENG J, et al. China national diabetes and metabolic disorders study group. Prevalence of diabetes among men and women in china ［J］. N Engl J Med, 2010, 362(12): 1090-1101.

［2］DAHL U, SJODIN A, SEMB H, et al. Cadherins regulate aggregation of pancreatic beta-cells in vivo ［J］. Development, 1996, 122: 2895-2902.

［3］HANAHAN D. Heritable formation of pancreatic beta-cell tumours in transgenic mice expressing recombinant insulin/simian virus 40 oncogenes ［J］. Nature, 1985, 315: 115-122.

［4］VASAVADA R C, CAVALIERE C, D’ERCOLE A J, et al. Overexpression of parathyroid hormone-related protein in the pancreatic islets of transgenic mice causes islet hyperplasia, hyperinsulinemia, and hypoglycemia ［J］. Biol Chem, 1996, 271: 1200-1208.

［5］HARA M, WANG X, KAWAMURA T, et al. Transgenic mice with green fluorescent protein-labeled pancreatic beta-cells ［J］. Am J Physiol Endocrinol Metab, 2003, 284: 177-183.

［6］HOTTA M, TASHIRO F, IKEGAMI H, et al. Pancreatic beta cell-specific expression of thioredoxin, an antioxidative and antiapoptotic protein, prevents autoimmune and streptozotocin-induced diabetes ［J］. Exp Med, 1998, 188: 1445-1451.

［7］KRAKOWSKI M L, KRITZIK M R, JONES E M, et al. Transgenic expression of epidermal growth factor and keratinocyte growth factor in beta-cells results in substantial morphological changes ［J］. J Endocrinol, 1999, 162: 167-175.

［8］MARTIN J, HUNT S L, DUBUS P, et al. Genetic rescue of Cdk4 null mice restores pancreatic beta-cell proliferation but not homeostatic cell number ［J］.Oncogene, 2003, 22: 5261-5269.

［9］SPURLOCK M E, GABLER N K. The development of porcine models of obesity and the metabolic syndrome ［J］. J Nutr, 2008, 138: 397-402.

［10］LONDRIGAN S L, BRADY J L, SUTHERLAND R M, et al. Evaluation of promoters for driving efficient transgene expression in neonatal porcine islets ［J］. Xenotransplantation, 2007,14: 119-125.

［11］袁小清，马向华，沈捷.核转录因子CHOP的研究进展［J］. 医学研究杂志，2008，8（37）: 15-18.

［12］周辉，张旭家.线粒体解偶联蛋白UCP2的研究进展［J］. 生命科学，2008，4（20）: 549-559.

［13］KHOI C, GAILLARD N E, TSAI M J. Beta2 and pancreatic islet development ［J］. En Rev, 2001, 56(1): 23-46.

［14］YANG K C, QI Z, YANAI G, et al. Cell coupling regulates Ins1, Pdx-1 and MafA to promote insulin secretion in mouse pancreatic beta cells ［J］. Process Biochem, 2011, 46: 1853-1860.

［15］MELLOUL D, MARSHAK S, CERASI E. Regulation of insulin gene transcription ［J］. Diabetologia March, 2002, 45(3): 309-326.

［16］BOAM D, CLARK A, DOCHERTY K. Positive and negative regulation of the insulin gene by multiple trans-acting factors［J］. Biol Chem, 1990, 265: 8285-8296.

［17］GERMAN M, RUTTER W J. The insulin gene promoter: a simplified nomenclature ［J］. Diabetes, 1995, 44(8): 1002-1004.

［18］WATANABE M, UMEYAMA K, KAWANO H, et al. The production of a diabetic mouse using constructs encoding porcine insulin promoter-driven mutant human hepatocyte nuclear factor-1alpha［J］. Reprod, 2007, 12(53): 189-200.

［19］MARJETA G, DAHLHOFF M, HERBACH N, et al. Specific transgene expression in mouse pancreatic β-cells under the control of the porcine insulin promoter ［J］. Mol Cell Endocrinol, 2010, 315(1-2): 219-224.

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Analysis of the Porcine Insulin Promoter and Verification of the Regulatory Genes Specific Expression in Pancreatic Tissue

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