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用于猪的强力霉素调节生长激素基因表达系统。

Doxycycline-regulated growth hormone gene expression system for swine.

作者信息

Jiang B C, Yu D B, Wang L J, Dong F L, Kaleri H A, Wang X G, Ally N, Li J, Liu H L

机构信息

Department of Animal Breeding & Genetics, College of Animal Science & Technology, Nanjing Agricultural University, Nanjing, China.

出版信息

Genet Mol Res. 2012 Aug 29;11(3):2946-57. doi: 10.4238/2012.July.10.5.

Abstract

Although growth hormone gene transgenic animals are much larger than normal animals, they manifest differences that have adverse effects on survival due to overexpression of growth hormone. We developed a stable pig embryonic fibroblast cell line expressing pig growth hormone (pGH) using the Tet-On system, with which we can conditionally manipulate expression of pGH in vivo. Inducible expression of pGH was achieved by combining reserve Tet-controlled transcriptional activator and tetracycline-responsive element in a single plasmid. The mRNA expression of pGH was significantly increased compared to the non-induced group by about 10-fold. The controlled secretion of pGH induced by doxycycline was further tested in stably transfected cells. We conclude that inducible GH expression can be achieved in pig embryonic fibroblasts.

摘要

尽管生长激素基因转基因动物比正常动物大得多,但由于生长激素的过度表达,它们表现出对生存有不利影响的差异。我们使用Tet-On系统开发了一种稳定表达猪生长激素(pGH)的猪胚胎成纤维细胞系,利用该系统我们可以在体内有条件地操纵pGH的表达。通过在单个质粒中组合储备型四环素控制转录激活因子和四环素反应元件,实现了pGH的诱导表达。与未诱导组相比,pGH的mRNA表达显著增加了约10倍。在稳定转染的细胞中进一步测试了强力霉素诱导的pGH的可控分泌。我们得出结论,猪胚胎成纤维细胞中可以实现诱导型GH表达。

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