[Expression and identification of F1 antigen of Y.pestis in Saccharomyces cerevisiae].
OBJECTIVE
To construct the recombinant vector containing F1 gene caf1 of Y.pestis and express it in Saccharomyces cerevisiae.


METHODS
F1 gene caf1 of Y.pestis was inserted into pHSS6-mTn-3xHA/lacZ plasmid to construct recombinant vector pHSS6-mTn-3xHA/lacZ-caf1. The recombinant plasmid was linearized with Not I and then transformed into yeast cells by acetate lithium (LiAc) method. Positive recombinants were selected with uracil-lack medium. The expressed F1 antigen was identified by SDS-PAGE and Western blot.


RESULTS
SDS-PAGE and Western blot analysis showed that F1 antigen was expressed in Saccharomyces cerevisiae.


CONCLUSIONS
The recombinant vector pHSS6-mTn-3xHA/lacZ-caf1 has been constructed and expressed successfully in Saccharomyces cerevisiae. These results lay the foundation for preparing gene vaccine of Y.pestis which could be taken via alimentary tract pathway.
Institute of Tumor Molecular Biology, General Hospital of Guangzhou Command, Guangzhou 510010, China.
Bin
B
Liu
Wen-ling
WL
Zheng
Hai
H
Deng
Wen-li
WL
Ma
D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology*; D001428:Bacterial Vaccines / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D015010:Yersinia pestis / Q000235:genetics / Q000276:immunology*
D000942:Antigens, Bacterial; D001428:Bacterial Vaccines