Expression of domain-3 envelop protein of Dengue virus type 2 in Escherichia coli
- Author:
Truong Uyen Ninh
- Publication Type:Journal Article
- Keywords:
Dengue virus type 2;
Eschirichia coli;
Evelop protein;
domain-3 encod4e;
Espression;
- MeSH:
Dengue Virus;
Escherichia coli;
Antigens
- From:Journal of Preventive Medicine
2005;15(5):138-144
- CountryViet Nam
- Language:Vietnamese
-
Abstract:
Envelope protein (E) of Dengue virus is an important antigen for immune response of Dengue virus, in which domain 3 (E3) encodes important specific epitopes of each serotype of the virus. It would be of a great value to express domain 3 and apply the recombinant proteins for development of the kit for DF/DHF diagnosis. The domain 3 of the gene coding for the envelope protein of dengue virus type 2 was amplified by PCR with primer pair that hangs BamHI and Xhol restriction sites at the 5 prime of the forward and reverse primer, respectively. The template for domain-3 amplification was the gene encoding preM-E protein that had been cloned and sequenced before. The amplified region of the gene was then inserted into the pET-TRX Fusion Expression Vector System with BamHI and Xhol restriction sites and transformed into E. coli BL21 DE3 Star. Recombinant protein D2E3 was expressed in a fusion form with Thioredoxin and His-Tag after induction with ImM IPTG. Recombinant D2E3 was purified with ProBond Nickel-Chelating Resin column (Invitrogen) and the purified recombinant D2E3 protein was used to check the reaction with the antibodies against native dengue antigens in the patient's sera by ELISA, Western blot and Dot blot. The results showed that, recombinant D2E3 protein of Dengue virus type 2 could react specifically with antibodies in the patient's sera.
