牛PrPc结构蛋白基因亚克隆及原核表达
Sub-cloning and Prokaryotic Expression of Bovine PrPc Structural Protein Gene
目的 制备PrPc重组结构蛋白，为研究与人和动物海绵状脑病发病有关的膜蛋白PrPc的生物学功用提供试验材料。方法 采用PCR扩增并克隆奶牛PrPc基因,经DNA序列分析后, 亚克隆至PET30a(+)表达质粒，优化条件进行目的蛋白表达。结果 PrPc基因序列与已发表的标准PrP基因序列相同。在蛋白原核表达系统中,标准PrPc基因可有效地表达完整的PrPc蛋白。Western blotting蛋白转印试验证实, 所表达的蛋白具有良好的免疫反应性。 结论 牛PrPc基因能在大肠杆菌蛋白表达系统中得到有效地表达。
Object To prepare recombinant PrPc structural protein, and this would be used for studying of biological function of Prion protein. Methods Bovine PrPc structural protein gene was amplified by PCR from genome DNA and cloned into PET30a(+) vector, protein was expressed after option of experiment condition. Results The results described here showed that the highest yield of recombinant protein expression induced by IPTG, under 37℃ for 3h. The expression protein and peptide antibody has good reaction by western blotting analysis. Conclusion The results indicated that we had successfully developed recombinant protein of bovine prion, which would be used for further structural biology study of prion.
李宁、郝永新、韩彩霞、杨建民、宁章勇、吴长德
分子生物学生物工程学
朊病毒奶牛PrPc表达
TSEPrPcProteinSub-cloningexpression
李宁,郝永新,韩彩霞,杨建民,宁章勇,吴长德.牛PrPc结构蛋白基因亚克隆及原核表达[EB/OL].(2005-12-28)[2025-08-02].http://www.paper.edu.cn/releasepaper/content/200512-721.点此复制
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