Construction and expression of HSV-2gD-Hsp70 fusion protein gene / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 914-918, 2006.
Article
in Chinese
| WPRIM
| ID: wpr-325450
ABSTRACT
To construct and express Hsp70-HSV2gD fusion protein. Genes of Hsp70 and HSV-2gD were subcloned into vectors pGEX-4T-1 respectively. After confirmed by DNA sequence analysis, the recombinant plasmids pGEX-4T-HSP-gD was transformed into E. coli DH5alpha and induced to express with IPTG. The expressed protein was characterized by SDS-PAGE and Western blot after purified. BALB/c mice were immunized with fusion proteins respectively via intra-m uscular injection. The proliferation of spleen lymphocytes, the level of y-IFN in culture and anti-HSV-2gD IgG antibody in serum was detected was detected. The expressed protein was analyzed by SDS-PAGE after induced with IPTG, which showed a new band with an apparent molecular mass corresponding to the predicted size (118 kD). Western Blotting analysis demonstrates that the purified Hsp70-HSV2gD fusion protein had specific binding activity. The stimulation indexes of spleen lymphocytes, the level of gamma-IFN in culture and anti-HSV-2gD IgG antibody in serum of GST-Hsp70-gD group was obviously higher than that of other groups (P < 0.05 respectively). The successful expression of the Hsp70-HSV2gD fusion protein, which can induce immune responses, laid a solid foundation for its further research.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Spleen
/
Blood
/
Recombinant Fusion Proteins
/
Immunoglobulin G
/
Gene Expression
/
Viral Envelope Proteins
/
Blotting, Western
/
Interferon-gamma
/
HSP70 Heat-Shock Proteins
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2006
Type:
Article
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