陕西科技大学学报(自然科学版)2007，Vol.25Issue(6)：1-6,6.
人源蛋白酶体α亚基6的酵母表面展示及表达优化
YEAST SURFACE DISPLAY OF HUMAN PROTEASOME SUBUNIT ALPHA 6 AND EXPRESSION ENHANCEMENT
摘要
Abstract
To construct the yeast display system of the human proteasome subunit alpha 6(hPSA6) for epitope analysis and mechanism investigation of ubiquitin-proteasome pathway,and enhance the display expression of hPSA6, the gene PSA6_HUMAN coding hPSA6 was cloned into a yeast-displaying expression vector, pICAS-H, which had been inserted a His. Tag marker for expression level detection. As probed with a his. Tag monoclonal antibody (Mab) and corresponding Mab, hPSA6 was detected functionally by flow cytometry and fluorescence microscopy analysis which confirmed that yeast-displaying recombinant hPSA6 with highly specific affinity was expressed efficiently after 24 h cultivation of recombinant yeast MT8-1/pICAS-H-PSA6. Induced by different concentration of initial glucose and casein acid in restrictive mediums, the displayed hPSA6 expressions were compared through immunofluorescence by using anti-His Mabs. Comparing with negative control, more than 70％ cells were induced to express hPSA6 in SA with 3％ casein acid initially. The 3％ glucose could make more than half cells express well and nearly 40％ cells were displaying hPSA6 successfully under the 2% initial glucose. Considering the glucose effect and the carbonization in sterilization period, 2％ glucose was more appropriate than 3％ in hPSA6 displaying.关键词
泛素-蛋白酶体途径/蛋白酶体α亚基6/酵母表面展示Key words
ubiquitin-proteasome pathway/ proteasome subunit alpha 6/ yeast surface display分类
生物学引用本文复制引用
唐语谦,林影,韩双艳,梁世中..人源蛋白酶体α亚基6的酵母表面展示及表达优化[J].陕西科技大学学报(自然科学版),2007,25(6):1-6,6.基金项目
国家科技攻关计划项目(2004BA711A20) （2004BA711A20）
