PMID- 14501197
OWN - NLM
STAT- MEDLINE
DCOM- 20031211
LR  - 20200324
IS  - 0920-8569 (Print)
IS  - 1572-994X (Electronic)
IS  - 0920-8569 (Linking)
VI  - 27
IP  - 2
DP  - 2003 Oct
TI  - High-level expression of the ORF6 gene of porcine reproductive and respiratory 
      syndrome virus (PRRSV) in Pichia pastoris.
PG  - 189-96
AB  - High-level expression of the ORF6 gene of porcine reproductive and respiratory 
      syndrome virus (PRRSV) has been proved very difficult. In this work, we cloned 
      and sequenced the ORF6 gene of PRRSV and found that it could not be expressed in 
      Pichia pastoris strain GS115. Then, the ORF6 gene was modified and synthesized 
      based on the codon bias, poly (A) signal of yeast expression system and secondary 
      structure of 5'-end mRNA of foreign gene. The modified gene was inserted into the 
      yeast expression vector pPICZalphaA, induced and expressed by the same methods. 
      The recombinant protein with a molecular mass of approximately 23 kDa was 
      screened by SDS-PAGE and identified by Western blot with convalescent sera of 
      animals infected with CH-1a strain of PRRSV. The results indicated that it was 
      similar to the native protein. The expression level of the recombinant protein 
      could attain 2.0 g/L. In the meanwhile, the optimal conditions for expression 
      were determined. It provides an additional means for studying the structural and 
      functional characteristics of PRRSV ORF6 gene.
FAU - Qian, Ping
AU  - Qian P
AD  - Laboratory of Animal Virology, College of Vet. Med. Science, Huazhong 
      Agricultural University, Wuhan, 430070, P.R. China.
FAU - Li, Xiangmin
AU  - Li X
FAU - Tong, Guangzhi
AU  - Tong G
FAU - Chen, Huanchun
AU  - Chen H
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Virus Genes
JT  - Virus genes
JID - 8803967
RN  - 0 (3' Untranslated Regions)
RN  - 0 (5' Untranslated Regions)
RN  - 0 (Codon)
RN  - 0 (RNA, Viral)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Viral Matrix Proteins)
RN  - Y4S76JWI15 (Methanol)
SB  - IM
MH  - 3' Untranslated Regions/genetics
MH  - 5' Untranslated Regions/genetics
MH  - Animals
MH  - Base Sequence
MH  - Blotting, Western
MH  - *Cloning, Molecular
MH  - Codon/genetics
MH  - Electrophoresis, Polyacrylamide Gel
MH  - *Gene Expression
MH  - Genetic Vectors
MH  - Methanol/metabolism
MH  - Molecular Sequence Data
MH  - Open Reading Frames/*genetics
MH  - Pichia/*genetics/growth & development/metabolism
MH  - Polymerase Chain Reaction
MH  - Porcine respiratory and reproductive syndrome virus/*genetics
MH  - RNA, Viral/genetics/isolation & purification
MH  - Recombinant Proteins/biosynthesis
MH  - Transcription, Genetic
MH  - Transformation, Genetic
MH  - Viral Matrix Proteins/biosynthesis/*genetics
PMC - PMC7089132
EDAT- 2003/09/23 05:00
MHDA- 2003/12/12 05:00
PMCR- 2020/03/23
CRDT- 2003/09/23 05:00
PHST- 2003/09/23 05:00 [pubmed]
PHST- 2003/12/12 05:00 [medline]
PHST- 2003/09/23 05:00 [entrez]
PHST- 2020/03/23 00:00 [pmc-release]
AID - 5142901 [pii]
AID - 10.1023/a:1025784712265 [doi]
PST - ppublish
SO  - Virus Genes. 2003 Oct;27(2):189-96. doi: 10.1023/a:1025784712265.