Cloning and Expression of Genes for Dengue Virus Type-2 Encoded Antigens for Rapid Diagnosis and Vaccine Development
Abstract
This Progress Report can be divided into five parts focusing on the overall goal of determining the complete sequence analysis of dengue virus genome, the expression of the cloned genes, especially the gene for the structural surface glycoprotein E in Escherichia Coli and characterization of the dengue viral antigens synthesized in the infected cell using the antibodies raised against the different polypeptide segments of the dengue viral polyprotein, expressed in E. coli. 1. Verification of the DNA sequence data previously generated for pVV1 and pVV9 clones, mapping in the region of dengue virus genome coding for nonstructural proteins. 2. Chemical synthesis of oligodeoxynucleotide primers, specific for the region encoded in pVV9, pVV17 and 3' end of the dengue virus genome. Verification of these primers for the specificity by hybridization with pVV1 and pVV9 clones. 3. Synthesis of cDNA copy of the dengue virus type 2 RNA in a reverse transcriptase catalyzed reaction, followed by double-stranded cDNA synthesis and transformation of E. coli JM83/pUC18 host/vector system. 4. Screening of the recombinant cDNA clones by restriction enzyme analysis and by Southern hybridization. 5. Synthesis and characterization of short peptide antigens, representing short hydrophilic segments of dengue viral proteins, the amino acid sequences of which were deduced by our DNA sequence data.
Document Details
- Document Type
- Technical Report
- Publication Date
- Nov 26, 1986
- Accession Number
- ADA203245
Entities
People
- Radha K. Padmanabhan
Organizations
- University of Kansas Medical Center