Preparation of Brucella canis Ghosts Vaccine

QIAN Jing;YIN Qiu-dan;BU Zhao-yang;CHEN Si;WANG Xing-long;WANG Xiu-ran;LANG Xu-long;WANG Xiao-xu;College of Animal Science and Technology,Jilin Agricultural University;Institute of Military Veterinary,Academy of Military Medical Sciences;Institute of Military Veterinary,Academy of Military Medical Sciences,Changchun 130122,China;Key Laboratory of Jilin Province for Zoonosis Prevention and Control;

Brucella canis;;bacterial ghost;;vaccine

Bacterial ghost technology is a novel method for preparing the inactivated vaccine.This nondenaturing inactivated method keeps many antigenic determinants which serves as a candidate vaccine to prevent pathogenic bacteria.This experiment took lysis Egene from phage PhiX174DNA by PCR and inserted Egene into pBV220vector,then cloned protein lysis component(PLC)from expression vector pBV220+E which we had constructed before,this PLC contained repressor protein cI857, lysis Egene and terminator sequence rrnbT1T2,as well inserted PLC into the broad-host shuttle vector pBBR1MCS-2.Finally,the broad-host lysis plasmid pBBR+PLC was electrical transformed into Brucella canis RM6/66.The results showed that the broad-host lysis plasmid had highly killing effect for Brucella canis RM6/66after 42 ℃induction and its lysis rate was 100%,meanwhile Brucella canis RM6/66ghosts were successfully generated.This study used bacterial ghost technology to prepare the Brucella canis ghost,which played an important role in the prevention of canine brucellosis,simultaneously provided a new strategy for human and animals' vaccine against brucellosis.