苦荞过敏原TBa和TBb基因的共表达及其包涵体复性的研究  

Coexpression of Tartary Buckwheat Allergen TBa and TBb and Renaturation of the Inclusion Body

作　　者：贺东亮[1] 张政[1] 任晓霞[1] 崔晓东[1] 李玉英[1] 王转花[1] 

机构地区：[1]化学生物学与分子工程教育部重点实验室,山西大学生物技术研究所,太原030006

出　　处：《中国农学通报》2009年第8期50-52,共3页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金(30870525;30671084);山西省自然科学基金(2007011077)

摘　　要：【研究目的】利用大肠杆菌共表达苦荞过敏蛋白的两个亚基,并对表达产物进行包涵体复性研究和免疫学活性分析;【方法】用已构建的表达质粒pET-28a-TBa和pET-32m-TBb,共转化大肠杆菌BL21(DE3)感受态,利用双抗生素筛选法,获得稳定遗传的双质粒转化子,经IPTG诱导,两个基因在同一宿主菌中共表达,在共表达产物复性过程中,两个亚基互为分子伴侣,相互促进了蛋白质的重新正确折叠;【结果】表达蛋白以包涵体的形式存在,ELISA检测表明:复性后的蛋白免疫学活性得到了提高;【结论】由此获得了有活性的蛋白质,并且建立了不相容双质粒共表达外源基因和包涵体复性的方法。[OBJECTIVE]To co-express the two subunits of Tartary Buckwheat Allergen in E.coli and made researchs on renaturation of inclusion body and check the immunologic characters of the production expressed, respectively; [METHOD] The two resuliting plasmids pET-28a-Tba and pET-32m-TBb were uesd to cotransform E.coli BL21（DE3） cell. After both ampicilln and kanamycin were presented in the selective medium and induction with IPTG, both TBa and TBb genes were coexpressed in E.coli. In the process of renaturation of coexpressed product, two subunits acted as molecular chaperones with each other, which promoted the refolding of protein TBa and TBb; [RESULT]Production expressed presentsed as inclusion body, and ELISA indicated that the renatured protein had the improvement on the immunological activity and gained the activated protein; [CONCLUSION] A new method for coexpression of proteins in E.coli containing two incompatible plasmids in which two different antibiotic resistant markers were included and inclusion body renaturation were also established.

关 键 词：苦荞 过敏蛋白 共表达 包涵体 复性 

分 类 号：Q78[生物学—分子生物学]