PMID- 22285974
OWN - NLM
STAT- MEDLINE
DCOM- 20120409
LR  - 20211021
IS  - 1879-0038 (Electronic)
IS  - 0378-1119 (Print)
IS  - 0378-1119 (Linking)
VI  - 496
IP  - 2
DP  - 2012 Apr 1
TI  - Analysis of the 5' untranslated region (5'UTR) of the alcohol oxidase 1 (AOX1) 
      gene in recombinant protein expression in Pichia pastoris.
PG  - 118-27
AB  - Pichia pastoris is a methylotrophic yeast that has been genetically engineered to 
      express over one thousand heterologous proteins valued for industrial, 
      pharmaceutical and basic research purposes. In most cases, the 5' untranslated 
      region (UTR) of the alcohol oxidase 1 (AOX1) gene is fused to the coding sequence 
      of the recombinant gene for protein expression in this yeast. Because the effect 
      of the AOX1 5'UTR on protein expression is not known, site-directed mutagenesis 
      was performed in order to decrease or increase the length of this region. Both of 
      these types of changes were shown to affect translational efficiency, not 
      transcript stability. While increasing the length of the 5'UTR clearly decreased 
      expression of a beta-galactosidase reporter in a proportional manner, a deletion 
      analysis demonstrated that the AOX1 5'UTR contains a complex mixture of both 
      positive and negative cis-acting elements, suggesting that the construction of a 
      synthetic 5'UTR optimized for a higher level of expression may be challenging.
CI  - Copyright A(c) 2012 Elsevier B.V. All rights reserved.
FAU - Staley, Chris A
AU  - Staley CA
AD  - Department of Biological Sciences, University of the Pacific, Stockton, CA 95211, 
      USA.
FAU - Huang, Amy
AU  - Huang A
FAU - Nattestad, Maria
AU  - Nattestad M
FAU - Oshiro, Kristin T
AU  - Oshiro KT
FAU - Ray, Laura E
AU  - Ray LE
FAU - Mulye, Tejas
AU  - Mulye T
FAU - Li, Zhiguo Harry
AU  - Li ZH
FAU - Le, Thu
AU  - Le T
FAU - Stephens, Justin J
AU  - Stephens JJ
FAU - Gomez, Seth R
AU  - Gomez SR
FAU - Moy, Allison D
AU  - Moy AD
FAU - Nguyen, Jackson C
AU  - Nguyen JC
FAU - Franz, Andreas H
AU  - Franz AH
FAU - Lin-Cereghino, Joan
AU  - Lin-Cereghino J
FAU - Lin-Cereghino, Geoff P
AU  - Lin-Cereghino GP
LA  - eng
GR  - R15 GM065882/GM/NIGMS NIH HHS/United States
GR  - R15 GM065882-03/GM/NIGMS NIH HHS/United States
GR  - GM65882-03/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20120125
PL  - Netherlands
TA  - Gene
JT  - Gene
JID - 7706761
RN  - 0 (5' Untranslated Regions)
RN  - 0 (Recombinant Proteins)
RN  - EC 1.1.- (Alcohol Oxidoreductases)
RN  - EC 1.1.3.13 (alcohol oxidase)
RN  - EC 3.2.1.23 (beta-Galactosidase)
SB  - IM
MH  - *5' Untranslated Regions
MH  - Alcohol Oxidoreductases/*genetics
MH  - Base Sequence
MH  - Cell-Free System
MH  - Gene Deletion
MH  - Gene Expression Profiling/methods
MH  - *Gene Expression Regulation, Fungal
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Mutation
MH  - Nucleic Acid Conformation
MH  - Pichia/*metabolism
MH  - Protein Biosynthesis
MH  - Real-Time Polymerase Chain Reaction/methods
MH  - Recombinant Proteins/metabolism
MH  - beta-Galactosidase/metabolism
PMC - PMC3464313
MID - NIHMS352105
EDAT- 2012/01/31 06:00
MHDA- 2012/04/10 06:00
PMCR- 2013/04/01
CRDT- 2012/01/31 06:00
PHST- 2011/09/14 00:00 [received]
PHST- 2011/12/31 00:00 [revised]
PHST- 2012/01/05 00:00 [accepted]
PHST- 2012/01/31 06:00 [entrez]
PHST- 2012/01/31 06:00 [pubmed]
PHST- 2012/04/10 06:00 [medline]
PHST- 2013/04/01 00:00 [pmc-release]
AID - S0378-1119(12)00021-2 [pii]
AID - 10.1016/j.gene.2012.01.006 [doi]
PST - ppublish
SO  - Gene. 2012 Apr 1;496(2):118-27. doi: 10.1016/j.gene.2012.01.006. Epub 2012 Jan 
      25.