Construction of a prokaryotic expression vector of human tau multi-epitope peptide and immunogenicity of the expressed product / 南方医科大学学报
Journal of Southern Medical University
;
(12): 185-188, 2012.
Article
in Chinese
| WPRIM
| ID: wpr-265666
ABSTRACT
<p><b>OBJECTIVE</b>To construct a prokaryotic expression vector of human tau multiepitope peptide for examining the immunogenicity of a TauP1/P2 DNA vaccine in mice using the expressed product.</p><p><b>METHODS</b>The coding sequence of Tau multiepitope peptide gene was amplified from the plasmid pVAX1-Tau by PCR and inserted into the prokaryotic expression vector pGEX-4T-2 to construct the recombinant plasmid pGEX-4T-2-TauP1/P2. The positive recombinants were transformed into E.coli BL21 cells, and the expression of fusion protein GST-TauP1/P2 was induced by IPTG and identified by SDS-PAGE. Mice was immunized with TauP1/P2 DNA vaccine and the production of the specific antibodies was detected by Dot-blot analysis using the purified fusion protein.</p><p><b>RESULTS</b>A gene fragment 300 bp in length was amplified. Enzyme digestion and DNA sequencing verified correct construction of the prokaryotic expression plasmid pGEX-4T-2-TauP1/P2. The expression of target fusion protein GST-TauP1/P2 was detected by SDS-PAGE. Specific antibodies against TauP1/P2 were detected in the serum of mice immunized with the DNA vaccine using GST-TauP1/P2 fusion protein.</p><p><b>CONCLUSION</b>The constructed prokaryotic expression plasmid of human Tau multiepitope peptide is capable of expressing the target fusion protein, which specifically recognizes the specific antibodies against TauP1/P2 in mice immunized with TauP1/P2 DNA vaccine.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Peptides
/
Recombinant Proteins
/
Tau Proteins
/
Vaccines, DNA
/
Allergy and Immunology
/
Escherichia coli
/
Genetic Vectors
/
Genetics
/
Metabolism
/
Mice, Inbred C57BL
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Journal of Southern Medical University
Year:
2012
Type:
Article
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