含35S启动子的粟酒裂殖酵母表达载体的构建及功能鉴定  

Construction and Identification of Schizosaccharomyces pombe Expression Vector Containing CaMV 35S Promoter

作　　者：王贺[1] 王丕武[2] 洪洋[1] 宋冰[2] 

机构地区：[1]吉林农业大学生命科学学院,长春130118 [2]吉林农业大学农学院,长春130118

出　　处：《生物技术通报》2011年第7期191-196,共6页Biotechnology Bulletin

基　　金：教育部博士点基金资助项目(20070193005)

摘　　要：利用PCR技术从植物表达载体pBI121上先后克隆出GUS基因和35S-GUS-NOS基因序列,分别构建粟酒裂殖酵母重组表达载体pESP-2-GUS和pESP-2-35S.GUS.NOS,并将它们转化到粟酒裂殖酵母菌体中。通过对不同液体培养基中两种转化子的不同启动子启动GUS基因表达产物的GUS染色分析与比较,证明连入的35S启动子能够替代pESP-2自身受维生素B1浓度调控的启动子,从而降低了培养基的成本,为今后工业上利用粟酒裂殖酵母发酵生产产品,提高经济效益,奠定了基础。Using PCR technology,we cloned GUS gene and 35S-GUS-NOS gene sequence from the plant expression vector pBI121,and constructed Schizosaccharomyces pombe＇s reorganization expression vector pESP-2-GUS and pESP-2-35S.GUS.NOS respectively,which were transmitted into the body of Schizosaccharomyces pombe.Through comparing and analyzing GUS staining,which is the product of GUS gene expression activated by promoters of two kinds of transforments in different liquid culture mediums,it proved that 35S promoter could replace pESP-2 promoter controlled by the concentration of Vitamin B1.It could decrease the cost of culture medium,and lay the foundation for using Schizosaccharomyces pombe to increase economic benefit in industry.

关 键 词：粟酒裂殖酵母 35S启动子 载体改造 GUS染色 

分 类 号：Q78[生物学—分子生物学]