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HBcAg基因在甲醇型酵母中的表达、产物纯化及其性质的初步研究
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作者 陈恒 黄亚红 +1 位作者 梁婉琪 张惟杰 《中国生物工程杂志》 CAS CSCD 2003年第9期85-90,共6页
为研究乙肝核心抗原蛋白 (HBcAg)在甲醇型酵母 (PichiaPastoris)中的表达和性质 ,用PCR方法将HBcAg基因 (L)克隆到P .Pastoris胞内表达载体pPIC3k中 ,并利用电击和同源重组法 ,将重组质粒pPIC3k L转化感受态的甲醇型GS1 1 5酵母菌株。... 为研究乙肝核心抗原蛋白 (HBcAg)在甲醇型酵母 (PichiaPastoris)中的表达和性质 ,用PCR方法将HBcAg基因 (L)克隆到P .Pastoris胞内表达载体pPIC3k中 ,并利用电击和同源重组法 ,将重组质粒pPIC3k L转化感受态的甲醇型GS1 1 5酵母菌株。经过筛选得到阳性P .Pastoris重组子。重组菌株经甲醇诱导培养 ,表达产物的Western印迹结果表明 ,HBcAg蛋白能在甲醇型酵母(PichiaPastoris)中诱导表达 ,产物为一 2 1 5kDa的蛋白。经蔗糖密度梯度超离心和CsCl密度梯度超离心纯化后 ,ELISA和密度测定结果表明重组HBcAg蛋白主要分布在密度为 1 2 5 76g ml和1 30 1 3g ml的 2个峰值处。电镜观察表明 ,该重组HBcAg蛋白能自主装配成大小不同的 2种颗粒 (即核心颗粒 ) ,大颗粒直径约 34nm左右 ,小颗粒直径约 30nm左右。同时 ,我们还观察到 ,该核心蛋白颗粒在体外可发生集聚现象。 展开更多
关键词 乙肝病毒 HBcAg基因 甲醇型酵母 表达 产物纯化 核心颗粒 抗原性
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Metabolomics Study in Methylotrophic Yeast: A Minireview
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作者 Jye Ping Fam Suriana Sabri +2 位作者 Syarul Nataqain Baharum Abu Bakar Salleh Siti Nurbaya Oslan 《Journal of Life Sciences》 2017年第1期11-18,共8页
Methylotrophic yeast has been used as a cost-effective and valuable host for expression of recombinant protein due to its unique methanol utilisation pathway. It has an AOX (alcohol oxidase) protein which has been c... Methylotrophic yeast has been used as a cost-effective and valuable host for expression of recombinant protein due to its unique methanol utilisation pathway. It has an AOX (alcohol oxidase) protein which has been characterised to be a strong and tightly methanol-inducible dependent promoter. Metabolomics is the systematic study and inclusive analysis of small molecules called metabolites in a biological system. Metabolomics plays an important part in connecting the phenotype and genotype gap because it magnifies the modifications in the proteome and provides a better phenotype representation of an organism. This quantitative study has provided a new perception on the metabolic burden derived from the overexpression of recombinant protein in methylotrophic yeast. In this review, we discuss the fundamental aspect of metabolomics in methylotrophic yeast followed by their latest developments. 展开更多
关键词 Methylotrophic yeast metabolomics methanol utilisation pathway.
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