[Construction of the recombinant secretion type BCG-Eg95 vaccine of Echinococcus granulosus].
OBJECTIVE
To construct the recombinant secretion type BCG-Eg95 vaccine of Echinococcus granulosus (rsBCG-Eg95).


METHODS
BCG-Ag85B signal sequence with 117 bp and Eg95 gene with 471 bp were amplified from the genome of BCG and pGEX-4T-Eg95 by PCR, respectively. BCG-Ag85B signal coding gene and Eg95 gene were cloned into E. coli-BCG shuttle-vector pMV261 to get the recombinant plasmid pSMEg95, which was confirmed by restriction endonuclease digestion, PCR amplification and gene sequencing. These recombinant plasmids were introduced into BCG by electroporation for the construction of rsBCG-Eg95 vaccine. The rsBCG-Eg95 positive clones were screened by Kan+ and identified by PCR amplification.


RESULTS
BCG-Ag85B signal sequence coding gene and Eg95 coding gene were successfully cloned into pMV261, which was confirmed by restriction endonuclease digestion, PCR amplification and sequencing of the plasmid pSMEg95. The plasmids were introduced into BCG and confirmed as the recombinant secreting BCG-Eg95 vaccine of E. granulosus (rsBCG-Eg95).


CONCLUSIONS
The recombinant secretion type BCG-Eg95 vaccine (rsBCG-Eg95) of E. granulosus with BCG-Ag85B signal sequence and Eg95 gene has been constructed.
He
Li-li
LL
Institute of Pathogenic Biology, Lanzhou University, Lanzhou 730000, China.
Jing
Tao
T
Zhu
Bing-dong
BD
Jia
Wan-zhong
WZ
D000818:Animals; D000947:Antigens, Helminth / Q000235:genetics / Q000276:immunology*; D001500:BCG Vaccine / Q000235:genetics / Q000276:immunology*; D048209:Echinococcus granulosus / Q000235:genetics / Q000276:immunology*; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology*; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology
D000947:Antigens, Helminth; D001500:BCG Vaccine; C446034:EG95 protein, Echinococcus granulosus; D015801:Helminth Proteins; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA