Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
21
pubmed:dateCreated
2008-10-28
pubmed:databankReference
pubmed:abstractText
We investigated the effect of codon optimization on the expression levels of heterologous proteins in Aspergillus oryzae, using the mite allergen Der f 7 as a model protein. A codon-optimized Der f 7 gene was synthesized according to the frequency of codon usage in A. oryzae by recursive PCR. Both native and optimized Der f 7 genes were expressed under the control of a high-level-expression promoter with their own signal peptides or in a fusion construct with A. oryzae glucoamylase (GlaA). Codon optimization markedly increased protein and mRNA production levels in both nonfused and GlaA-fused Der f 7 constructs. For constructs with native codons, analysis by 3' rapid amplification of cDNA ends revealed that poly(A) tracts tended to be added within the coding region, producing aberrant mRNAs that lack a termination codon. Insertion of a termination codon between the carrier GlaA and native Der f 7 proteins in the GlaA fusion construct resulted in increases in mRNA and secreted-carrier-GlaA levels. These results suggested that mRNAs without a termination codon as a result of premature polyadenylation are degraded, possibly through the nonstop mRNA decay pathway. We suggest that codon optimization in A. oryzae results in elimination of cryptic polyadenylation signals in native Der f 7, thereby circumventing the production of truncated transcripts and resulting in an increase in steady-state mRNA levels.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-10361683, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-10570197, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-10981683, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-11133486, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-11167959, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-11812218, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-11910109, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-11910110, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-11937640, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-11943375, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-12100052, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-12356476, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-12683242, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-12783481, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-1287665, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-1368682, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-1368725, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-14672702, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-1480491, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-15165245, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-15221230, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-15245907, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-15302836, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-15965016, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-16131587, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-16213112, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-16233305, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-16372000, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-16372009, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-16372010, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-16384792, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-16403645, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-17225093, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-1730842, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-17766461, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-5432063, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-6175758, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-7579660, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-7958922, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-8556554, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-8787393, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-9023927, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-9035405, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-9584084, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-9634791, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-9655346, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-9701599, http://linkedlifedata.com/resource/pubmed/commentcorrection/18791013-9701600
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
1098-5336
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
74
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
6538-46
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
2008
pubmed:articleTitle
Codon optimization increases steady-state mRNA levels in Aspergillus oryzae heterologous gene expression.
pubmed:affiliation
Department of Bioindustrial Informatics and Genomics, Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai 981-8555, Japan.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't