TYRAP酵母双杂交载体的构建

目的:肿瘤坏死因子(Tumor necrosis factor,TNF)是一种具有广泛生物学效应的细胞因子,在机体炎症和免疫防御反应中起着重要作用,其中包括运动刺激所产生的机体反应.一种新型蛋白TTRAP被认为在TNF信号通路中有可能起重要作用,为揭示其作用以及寻找与其作用的互作蛋白,构建了TTRAP酵母双杂交系统的"诱饵"裁体.方法:对含有人TTRAP全部序列的表达栽体pGEX-4T-1-TTRAP进行PCR扩增,酶切后将TTRAP基因连接到酵母双杂交系统的pLexA质粒上,得到"诱饵"(Bait)-pLexA-TTRAP,导入大肠杆菌扩增,筛选得到阳性克隆并提取重组质粒.结果:测序结果表明克隆正确.结论:成功构建pLexA-TTRAP酵母双杂交载体,为寻找TTRAP的互作蛋白打下基础.     

Construction of the yeast whole-cell Rhizopus oryzae lipase biocatalyst with high activity

Surface display is effectively utilized to construct a whole-cell biocatalyst. Codon optimization has been proven to be effective in maximizing production of heterologous proteins in yeast. Here, the cDNA sequence of Rhizopus oryzae lipase (ROL) was optimized and synthesized according to the codon bias of Saccharomyces cerevisiae, and based on the Saccharomyces cerevisiae cell surface display system with α-agglutinin as an anchor, recombinant yeast displaying fully codon-optimized ROL with high activity was successfully constructed. Compared with the wild-type ROL-displaying yeast, the activity of the codon-optimized ROL yeast whole-cell biocatalyst (25 U/g dried cells) was 12.8-fold higher in a hydrolysis reaction using p-nitrophenyl palmitate (pNPP) as the substrate. To our knowledge, this was the first attempt to combine the techniques of yeast surface display and codon optimization for whole-cell biocatalyst construction. Consequently, the yeast whole-cell ROL biocatalyst was constructed with high activity. The optimum pH and temperature for the yeast whole-cell ROL biocatalyst were pH 7.0 and 40 °C. Furthermore, this whole-cell biocatalyst was applied to the hydrolysis of tributyrin and the resulted conversion of butyric acid reached 96.91% after 144 h.     

糖蜜酵母废水处理技术研究进展

文章分析了酵母工业废水的特点,酵母生产工艺,酵母行业的趋势;综述了厌氧处理、厌氧-好氧两级处理、厌氧-好氧-缺氧(A2/O)反硝化处理、化学混凝法、氧化法、膜分离技术、农灌法、蒸发浓缩法、喷雾干燥、循环利用、生物流化床耦合工艺等技术。     

Immunomodulatory function of orally administered thymosin α1

INTRODUCTION The thymus is a vital immune organ and plays a very important role in the development and mainte- nance of the lymph system. The extract of animal thymus has been used in clinical practice as an ad- junct treatment in patients with carcinoma, viral in- fection and immunodeficiency, but the extract con- taining miscellaneous animal proteins may easily lead to allergy in patients. Thymosin α1 (Tα1) is the most potent ingredient in thymosin and the biological ac- tivity of pur…     

中国小麦花叶病毒运动蛋白基因的克隆及其表达

应用RT-PCR技术从小麦病叶中扩增得到中国小麦花叶病毒(CWMV)的运动蛋白(MP)基因,在大肠杆菌BL21(DE3)plysS中得到大量表达,并以含表达产物的凝胶为抗原,免疫小鼠制备特异性抗血清,同时将MP基因重组到质粒pGBKT7中,电击转化酵母Y187菌株。Westernblot分析表明MP基因在酵母体内能以融合蛋白的形式正确表达,并具有免疫活性,可作为酵母双杂交系统中的“诱饵”质粒。