PMID- 15249057
OWN - NLM
STAT- MEDLINE
DCOM- 20050131
LR  - 20061115
IS  - 1046-5928 (Print)
IS  - 1046-5928 (Linking)
VI  - 36
IP  - 2
DP  - 2004 Aug
TI  - Systematic optimization of active protein expression using GFP as a folding 
      reporter.
PG  - 327-32
AB  - Many recombinant proteins have been used as drugs; however, human proteins 
      expressed using heterologous hosts are often insoluble. To obtain correctly 
      folded active proteins, many optimizations of expression have been attempted but 
      usually are found to be applicable only for specific targets. Interleukin-18 
      (IL-18) has a key role in many severe disorders including autoimmune diseases, 
      and therapeutic approaches using IL-18 have been reported. However, production of 
      IL-18 in Escherichia coli resulted in extensive inclusion body formation and 
      previous conventional screenings of expression conditions could obtain only a 
      condition with a low yield. To address the problem, we applied a folding reporter 
      system using green fluorescent protein (GFP) for screening of the expression 
      conditions for hIL-18. The established system efficiently screened many 
      conditions, and optimized conditions for the expression of hIL-18 significantly 
      enhanced the final yield of the active protein. Systematic screening using a GFP 
      reporter system could be applied for the production of other proteins and in 
      other organisms.
FAU - Omoya, Kentaro
AU  - Omoya K
AD  - Department of Pediatrics, Graduate School of Medicine, Gifu University, Yanagido 
      1-1, Gifu, Gifu 501-1194, Japan.
FAU - Kato, Zenichiro
AU  - Kato Z
FAU - Matsukuma, Eiji
AU  - Matsukuma E
FAU - Li, Ailian
AU  - Li A
FAU - Hashimoto, Kazuyuki
AU  - Hashimoto K
FAU - Yamamoto, Yutaka
AU  - Yamamoto Y
FAU - Ohnishi, Hidenori
AU  - Ohnishi H
FAU - Kondo, Naomi
AU  - Kondo N
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Protein Expr Purif
JT  - Protein expression and purification
JID - 9101496
RN  - 0 (Interleukin-18)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - EC 2.5.1.18 (Glutathione Transferase)
SB  - IM
MH  - Escherichia coli/chemistry/genetics
MH  - Gene Expression
MH  - Genes, Reporter/genetics
MH  - Glutathione Transferase/*chemistry/genetics/isolation & purification
MH  - Green Fluorescent Proteins/*chemistry/genetics/isolation & purification
MH  - Humans
MH  - Inclusion Bodies/chemistry
MH  - Interleukin-18/*chemistry/genetics/isolation & purification
MH  - *Protein Folding
MH  - Recombinant Fusion Proteins/chemistry/genetics/isolation & purification
EDAT- 2004/07/14 05:00
MHDA- 2005/02/03 09:00
CRDT- 2004/07/14 05:00
PHST- 2004/04/01 00:00 [received]
PHST- 2004/04/27 00:00 [revised]
PHST- 2004/07/14 05:00 [pubmed]
PHST- 2005/02/03 09:00 [medline]
PHST- 2004/07/14 05:00 [entrez]
AID - S1046592804001524 [pii]
AID - 10.1016/j.pep.2004.04.023 [doi]
PST - ppublish
SO  - Protein Expr Purif. 2004 Aug;36(2):327-32. doi: 10.1016/j.pep.2004.04.023.