title: Enhancement of recombinant glucoamylase expression by introducing yeast GAL7 mRNA termination sequence.
abstract: Glucoamylase gene (STA1) of Saccharomyces diastaticus was expressed in recombinant Saccharomyces cerevisiae systems. The yeast, GAL7 mRNA termination sequence, was introduced in the 3' noncoding region of the STA1 structural gene which was under the control of the SUC2 promoter and STA1 secretion signal sequence. This plasmid was named YEpSSG7 and was introduced into yeast S. cerevisiae MMY2 to construct recombinant S. cerevisiae MMY2SSG7. The GAL7 mRNA termination sequence enhanced the glucoamylase expression level by 3-5 times depending on the culture conditions compared to the result from the strain S. cerevisiae MMY2SUCSTA which did not contain the GAL7 mRNA termination sequence. Such an enhancement was not due to plasmid stability or plasmid copy number effects. Such an enhancement was primarily due to the fact that GAL7 mRNA termination sequence stabilized the STA1 mRNA 3' end.
authors.affiliation: Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University, South Korea.
authors.forename: K M
authors.initials: KM
authors.lastname: Cho
authors.forename: H J
authors.initials: HJ
authors.lastname: Cha
authors.forename: Y J
authors.initials: YJ
authors.lastname: Yoo
authors.forename: J H
authors.initials: JH
authors.lastname: Seo
mesh_terms: D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D005087:Glucan 1,4-alpha-Glucosidase / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012331:RNA, Fungal / Q000737:chemistry / Q000235:genetics; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012440:Saccharomyces / Q000201:enzymology / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013172:Spores, Fungal / Q000201:enzymology; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic / Q000235:genetics; D014170:Transformation, Genetic
chemical_list: D012331:RNA, Fungal; D012333:RNA, Messenger; D011994:Recombinant Proteins; D015246:Deoxyribonuclease EcoRI; D015247:Deoxyribonuclease HindIII; D005087:Glucan 1,4-alpha-Glucosidase