The genc CPA and ETX encoding α and e toxins were amplified from the standard strain C60-2 of type D Clostridium perfingens by PCR, and then they were inserted into pet-Duct-1 vecto, respectively for the construction...The genc CPA and ETX encoding α and e toxins were amplified from the standard strain C60-2 of type D Clostridium perfingens by PCR, and then they were inserted into pet-Duct-1 vecto, respectively for the construction of co-expression plasmid pETDoet-1-CPA-ETX. The co-expression plasmid was transformed into BL21 （DE3） competent cells, at and e toxins proteins were induced by IPTG before analysis by SDS-PAGE. As a result, both of α and e toxins were detected at 43 and 34 KU by western-blot. Mice immunized with the co-expressed α and e toxins proteins produced high titers of neutralizing antibodies in the serum, which protected the mice against the attack of type D C. perfringens culture filtrate. In addition, mice immunized with the produced co-expressed α and e toxins proteins showed significantly higher surviving rate than with ε toxins protein alone when infected with culture filtrate of type D C. perfringens . These results indicated that co-expression of α and e toxins proteins could be used as a new method to prepare vaccines against the pathogens of multiple serotypes.展开更多
基金Supported by the Basic Scientific Research Project of Nonprofit Central Research Institutions(1610322013028)
文摘The genc CPA and ETX encoding α and e toxins were amplified from the standard strain C60-2 of type D Clostridium perfingens by PCR, and then they were inserted into pet-Duct-1 vecto, respectively for the construction of co-expression plasmid pETDoet-1-CPA-ETX. The co-expression plasmid was transformed into BL21 （DE3） competent cells, at and e toxins proteins were induced by IPTG before analysis by SDS-PAGE. As a result, both of α and e toxins were detected at 43 and 34 KU by western-blot. Mice immunized with the co-expressed α and e toxins proteins produced high titers of neutralizing antibodies in the serum, which protected the mice against the attack of type D C. perfringens culture filtrate. In addition, mice immunized with the produced co-expressed α and e toxins proteins showed significantly higher surviving rate than with ε toxins protein alone when infected with culture filtrate of type D C. perfringens . These results indicated that co-expression of α and e toxins proteins could be used as a new method to prepare vaccines against the pathogens of multiple serotypes.
