摘要
利用已克隆的BALB/c小鼠Doppel蛋白基因,将其成熟蛋白编码区亚克隆至表达载体pGEX-6P-1上,构建重组表达质粒pGEX-6P-1-PRND。将重组蛋白表达载体转入E.coli BL21(DE3)中诱导表达,SDS-PAGE及Western blot分析结果表明,BALB/c小鼠Doppel蛋白在大肠杆菌中以包涵体形式高效表达。对表达的蛋白进行提取和纯化并免疫新西兰大白兔制备兔抗血清,初步建立了小鼠睾丸Doppel蛋白组织学分布的免疫组织化学方法。为进一步研究Doppel的结构、功能及其在TSEs发生发展中的作用提供基础材料和数据。
Using the cloned Doppel protein gene of BALB/c mouse,the recombinant prokaryotic expression plasmid pGEX-6P-1-PRND was constructed by subcloning the maturation protein gene encoding sequence into the vector pGEX-6P-1.The recombinant plasmid pGEX-6P-1-PRND was transformed into E.coli BL21（DE3） and induced to express.SDS-PAGE and Western blot results showed that the Doppel protein of BALB/c mouse was highly expressed in the form of inclusion body.The New Zealand White rabbits were immuned by the purified expression protein to produce immunol sera.The method of immunohistochemistry was primarily established to detect the Doppel distribution of testis in BALB/c mouse.The results provide the basic data and materials for the further study of structure and function of the protein and its role in transmissible spongiform encephalopathies.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2011年第12期1777-1780,共4页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(30700603)
广东省自然科学基金资助项目(7300744)
