PMID- 30537227
OWN - NLM
STAT- MEDLINE
DCOM- 20190819
LR  - 20231213
IS  - 1097-0061 (Electronic)
IS  - 0749-503X (Linking)
VI  - 36
IP  - 5
DP  - 2019 May
TI  - YHp as a highly stable, hyper-copy, hyper-expression plasmid constructed using a 
      full 2-mum circle sequence in cir(0) strains of Saccharomyces cerevisiae.
PG  - 249-257
LID - 10.1002/yea.3371 [doi]
AB  - In the yeast Saccharomyces cerevisiae, the yeast episomal plasmid (YEp), 
      containing a partial sequence from a natural 2-mum plasmid, has been frequently 
      used to induce high levels of gene expression. In this study, we used Japanese 
      sake yeast natural cir(0) strain as a host for constructing an entire 2-mum 
      plasmid with an expression construct using the three-fragment gap-repair method 
      without Escherichia coli manipulation. The 2-mum plasmid contains two long 
      inverted repeats, which is problematic for the amplification by polymerase chain 
      reaction. Therefore, we amplified it by dividing into two fragments, each 
      containing a single repeat together with an overlapping sequence for homologous 
      recombination. TDH3 promoter-driven yEmRFP (TDH3p-yEmRFP) and the URA3 were used 
      as a reporter gene and a selection marker, respectively, and inserted at the 3' 
      end of the RAF1 gene on the 2-mum plasmid. The three fragments were combined and 
      used for the transformation of sake yeast cir(0) ura3(-) strain. The resulting 
      transformant colonies showed a red or purple coloration, which was significantly 
      stronger than that of the cells transformed with YEp-TDH3p-yEmRFP. The 2-mum 
      transformants were cultured in YPD medium and observed by fluorescence 
      microscopy. Almost all cells showed strong fluorescence, suggesting that the 
      plasmid was preserved during nonselective culture conditions. The constructed 
      plasmid maintained a high copy state similar to that of the natural 2-mum plasmid, 
      and the red fluorescent protein expression was 54 fold compared with the 
      chromosomal integrant. This vector is named YHp, the Yeast Hyper expression 
      plasmid.
CI  - (c) 2018 John Wiley & Sons, Ltd.
FAU - Misumi, Yukie
AU  - Misumi Y
AD  - Department of Applied Chemistry, Graduate School of Sciences and Technology for 
      Innovation, Yamaguchi University, Ube, Japan.
FAU - Nishioka, Satoko
AU  - Nishioka S
AD  - Department of Applied Chemistry, Graduate School of Sciences and Technology for 
      Innovation, Yamaguchi University, Ube, Japan.
FAU - Fukuda, Akira
AU  - Fukuda A
AD  - JXTG Nippon Oil & Energy Corporation, Yokohama, Japan.
FAU - Uemura, Takeshi
AU  - Uemura T
AD  - JXTG Nippon Oil & Energy Corporation, Yokohama, Japan.
FAU - Nakamura, Mikiko
AU  - Nakamura M
AD  - Department of Applied Chemistry, Graduate School of Sciences and Technology for 
      Innovation, Yamaguchi University, Ube, Japan.
FAU - Hoshida, Hisashi
AU  - Hoshida H
AD  - Department of Applied Chemistry, Graduate School of Sciences and Technology for 
      Innovation, Yamaguchi University, Ube, Japan.
FAU - Akada, Rinji
AU  - Akada R
AUID- ORCID: 0000-0001-9044-9917
AD  - Department of Applied Chemistry, Graduate School of Sciences and Technology for 
      Innovation, Yamaguchi University, Ube, Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190124
PL  - England
TA  - Yeast
JT  - Yeast (Chichester, England)
JID - 8607637
RN  - 0 (DNA, Fungal)
RN  - 0 (Luminescent Proteins)
MH  - Cloning, Molecular
MH  - DNA, Fungal/genetics
MH  - Fermented Foods/microbiology
MH  - Gene Expression
MH  - *Genes, Fungal
MH  - Luminescent Proteins/genetics
MH  - *Nucleic Acid Amplification Techniques
MH  - Plasmids/*genetics
MH  - Recombination, Genetic
MH  - Saccharomyces cerevisiae/*genetics
MH  - Red Fluorescent Protein
OTO - NOTNLM
OT  - 2-mum plasmid
OT  - YEp
OT  - gap-repair cloning
OT  - homologous recombination
OT  - recombinant DNA
OT  - sake yeast
EDAT- 2018/12/12 06:00
MHDA- 2019/08/20 06:00
CRDT- 2018/12/12 06:00
PHST- 2018/10/10 00:00 [received]
PHST- 2018/11/20 00:00 [revised]
PHST- 2018/11/24 00:00 [accepted]
PHST- 2018/12/12 06:00 [pubmed]
PHST- 2019/08/20 06:00 [medline]
PHST- 2018/12/12 06:00 [entrez]
AID - 10.1002/yea.3371 [doi]
PST - ppublish
SO  - Yeast. 2019 May;36(5):249-257. doi: 10.1002/yea.3371. Epub 2019 Jan 24.