Construction and expression of recombinant plasmid to initiate polarization and activation of regulatory T cells.
- Author:
Qian ZHANG
1
;
Gong-Jian QI
;
Dang-Yang WANG
;
Jian-Jun ZHANG
;
Chong CHEN
;
Ling-Yu ZENG
;
Kai-Lin XU
Author Information
1. Department of Hematology, Xuzhou Medical College Affiliated Hospital, Jiangsu Province, China.
- Publication Type:Journal Article
- MeSH:
Cloning, Molecular;
Escherichia coli;
metabolism;
Genetic Vectors;
Humans;
Lymphocyte Activation;
Plasmids;
genetics;
Recombinant Proteins;
genetics;
metabolism;
T-Lymphocytes, Regulatory;
cytology;
metabolism;
Transforming Growth Factor beta1;
genetics;
metabolism
- From:
Journal of Experimental Hematology
2011;19(5):1250-1254
- CountryChina
- Language:Chinese
-
Abstract:
In order to construct a recombinant plasmid initiating the polarization and activation of the regulatory T cells (Treg), the fragments of hTGF-β1 and C2-C4 of gp120 amplified by RT-PCR and cloned into pCR2.1 vector respectively. hTGF-β1 and C2-C4 DNA fragments were obtained, then sub-cloned to generate the prokaryotic expression vector named pET-28a/C2-C4-Linker- hTGF-β1. The expression of recombinant protein was induced by IPTG (0.1 mmol/L) for 6 hours. The results showed that the fragments of hTGF-β1 and C2-C4 were amplified and cloned into pCR2.1, the prokaryotic expression vector pET-28a/C2-C4-Linker- hTGF-β1 was constructed successfully. The recombinant protein was expressed as inclusion body after being induced by IPTG. It is concluded that this recombinant protein can initiate the polarization and activation of Treg cells, indicating the engineering E.coli strain is successfully obtained.
