PMID- 15969006
OWN - NLM
STAT- MEDLINE
DCOM- 20100429
LR  - 20201209
IS  - 1000-3061 (Print)
IS  - 1000-3061 (Linking)
VI  - 19
IP  - 3
DP  - 2003 May
TI  - [Construction of MAGE-3 prokaryotic expression plasmid and its expression in 
      Escherichia coli].
PG  - 277-80
AB  - To express the GST-MAGE-3 protein in E. coli, and investigate the antitumor 
      immune responses induced by Dendritic cells (DCs) pulsed with GST-MAGE-3 protein, 
      the recombinant expression plasmid pGEX-MAGE-3 was constructed by ligating MAGE-3 
      gene, which was amplified by RT-PCR and confirmed by sequencing, and the 
      pGEX-4T-2 vector. The recombinant plasmid was transformed into BL-21 E. coli. The 
      expression of GST-MAGE-3 was induced with IPTG. The GST-MAGE-3 protein expressed 
      as high as 32% of the total cellular protein. After purification with Glutathione 
      Sepharose 4B, the purity of the protein was more than 90%, and 3mg GST-MAGE-3 was 
      obtained from 100 mL BL-21 lysate. Dendritic cells from gastric carcinoma 
      patients were pulsed with GST-MAGE-3 protein, and these DCs were used to 
      stimulate the autologous T. lymphocytes. After 7 days, the T. lymphocytes 
      cocultured with DCs pulsed with GST-MAGE-3 antigen exhibited specific 
      cytotoxicity against MAGE-3 positive SGC-7901 cells. It is concluded that the 
      GST-MAGE-3 protein are able to present antigen to T. lymphocytes, activate 
      antigen-specific CTLs and induce special antitumor immune responses in vitro. Our 
      results lay the groundwork for further research of the MAGE-3 vaccine.
FAU - Sun, Xiao-Dong
AU  - Sun XD
AD  - Department of General Surgery, Sir Run Run Shaw Hospital, Zhejiang University, 
      Hangzhou 310016, China.
FAU - Wu, Jin-Min
AU  - Wu JM
FAU - Liu, Xing-E
AU  - Liu XE
LA  - chi
PT  - Journal Article
PL  - China
TA  - Sheng Wu Gong Cheng Xue Bao
JT  - Sheng wu gong cheng xue bao = Chinese journal of biotechnology
JID - 9426463
RN  - 0 (Antigens, Neoplasm)
RN  - 0 (MAGEA3 protein, human)
RN  - 0 (Neoplasm Proteins)
SB  - IM
MH  - Antigens, Neoplasm/genetics/isolation & purification/*metabolism/pharmacology
MH  - Blotting, Western
MH  - Cell Line, Tumor
MH  - Cells, Cultured
MH  - Dendritic Cells/drug effects/immunology
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Escherichia coli/genetics/*metabolism
MH  - Genetic Vectors/*genetics
MH  - Humans
MH  - Neoplasm Proteins/genetics/isolation & purification/*metabolism/pharmacology
MH  - Plasmids/*genetics
MH  - Stomach Neoplasms/immunology
MH  - T-Lymphocytes/immunology
EDAT- 2005/06/23 09:00
MHDA- 2010/04/30 06:00
CRDT- 2005/06/23 09:00
PHST- 2005/06/23 09:00 [pubmed]
PHST- 2010/04/30 06:00 [medline]
PHST- 2005/06/23 09:00 [entrez]
PST - ppublish
SO  - Sheng Wu Gong Cheng Xue Bao. 2003 May;19(3):277-80.