一种快速筛选酵母高表达菌株的新方法  

A Novel Method for Rapid Screening the High-level Expressing Pichia pastoris Transformants

作　　者：冯晶晶[1,2] 赵媛媛[1] 杨梅[1,3] 吕茂民[1] 

机构地区：[1]军事医学科学院野战输血研究所,北京100850 [2]吉林农业大学生物技术学院,长春130021 [3]扬州大学生物科学与技术学院,扬州225009

出　　处：《生物技术通报》2008年第6期168-169,174,共3页Biotechnology Bulletin

摘　　要：为建立一种快速筛选酵母高表达菌株的方法,将生长在固体培养基上的新转化的表达菌落原位转移到醋酸纤维素薄膜上,再利用硝酸纤维素薄膜原位捕获醋酸纤维素薄膜上的蛋白,然后用免疫印迹的方法检测原位转印到硝酸纤维素膜上菌落分泌蛋白,根据免疫印迹的强弱进行高表达菌株的筛选。结果显示,经过菌落原位转印和免疫印迹显色,显色信号强的菌落表达量相对较高。因此,双膜法原位检测是一种快速、高效的筛选酵母高表达菌株的方法。In order to establish a method for rapid screening the high-level expressing transformants,the transformants on the solid medium were transferred onto cellulose acetate filter,and then the secreted protein of transformants on cellulose acetate filters were captured by nitrocellulose filter in site.The immunoblotting was performed using corresponding antibody,and the high-level expressing transformants can be selected according to immunoblotting figure.result indicated that according to immunoblotting coloration dot,the high-level expressing transformants was selected.The darker dot was high-level transformants expressed.Therefore,the immunological based double filters screening method is a rapid way to detect high-level expressing transformants.

关 键 词：酵母 原位双膜法 筛选 

分 类 号：Q78[生物学—分子生物学] TQ920.1[轻工技术与工程—发酵工程]