摘要
用PCR合成的瑞氏木霉 (T .reesei) β 内切葡聚糖酶Ⅰ (EGⅠ )的cDNA ,构建了由酵母醇脱氢酶 (ADH1 )启动子和终止子引导表达、β 内切葡聚糖酶自身信号肽序列引导分泌、由酵母rDNA序列引导同源整合的酵母YIP型 β 内切葡聚糖酶表达分泌质粒pA1 5 PET。采用pA1 5 PET与酵母YEP型G41 8抗性表达质粒的共转化 ,将EGⅠ表达单元整合到已整合有α 乙酰乳酸脱羧酶 (α ALDC)表达单元的啤酒酵母工程菌BE971 1的染色体rDNA序列中 ,获得同时表达胞内α ALDC和胞外β
An integration plasmid pA15-PET for expression and secretion of β-Endoglucana se Ⅰ (EGⅠ) in yeast was constructed by insertion of EGⅠcDNA between yeas t alcohol dehydrogenase promoter and terminator region.The plasmid contained par t of yeast rDNA sequence,which was used as a homologous fragment for integration . The EGⅠcDNA was introduced into an engineered brewing yeast BE9711 containing α-acetolactate decarboxylase (α-ALDC) encoding gene and integrated onto its rDNA sequence of chromosomal DNA by co-transformation of pA15-PET and a YEP t ype plasmid pA15TXR carrying G418 resistance. The stable engineered brewing yea st expressing intracellulase α-ALDC and extracellular EGⅠsimultaneously were obtained.
出处
《微生物学报》
CAS
CSCD
北大核心
2003年第5期586-591,共6页
Acta Microbiologica Sinica
