犬布鲁氏菌菌壳疫苗的制备简

菌壳技术是一种新型的灭活疫苗制备方法，通过非变性的灭活方式保存细菌表面多个抗原表位，所制备的菌壳可作为预防细菌病的理想疫苗。本试验从噬菌体PhiX174 DNA钓取裂解E基因，连接至温控原核表达载体pBV220，通过PCR在所构建的pBV220+E上扩增出蛋白裂解部件（protein lysis component,PLC），该部件包含阻遏蛋白cI857、溶菌E基因及终止序列rrnbT1T2，然后将其克隆至广宿主表达载体pBBR1MCS-2中，最终将构建的广宿主裂解质粒pBBR+PLC电转入犬布鲁氏菌RM6/66中。试验结果表明，经42 ℃诱导后，广宿主裂解质粒对犬布鲁氏菌RM6/66的裂解率达100%，成功制备了犬布鲁氏菌RM6/66菌壳疫苗。本试验通过菌壳技术制备的犬布鲁氏菌疫苗对预防宠物犬布鲁氏菌病起到重要作用，同时也对人兽布鲁氏菌疫苗的研制提供新策略。

Preparation of Brucella canis Ghosts 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 E gene from phage PhiX174 DNA by PCR and inserted E gene into pBV220 vector, then cloned protein lysis component (PLC) from expression vector pBV220+E which we had constructed before, this PLC contained repressor protein cI857, lysis E gene 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 RM6/66 after 42 ℃ induction and its lysis rate was 100%, meanwhile Brucella canis RM6/66 ghosts 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.