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Cloning and expression of the gene encoding CS3 fimbriae antigen of enterotoxigenic Escherichia coli



Cloning and expression of the gene encoding CS3 fimbriae antigen of enterotoxigenic Escherichia coli



Zhonghua Weishengquxue He Mianyixue Zazhi 14(2): 84-88



The results of Southern blot revealed that the CS3 antigen was coded by a large plasmid (60MD) isolated from E44815 strain, and the DNA fragment containing the gene encoding CS3 fimbriae was about 5.0 kb when the plasmid was digested by Hind III. Limited gene bank was constructed by recovering all the DNA fragments of about 5.0 kb after the plasmid was digested and inserting it into the Hind III site of pBR322. Through screening, we obtained two kinds of positive recombinant plasmids harboring the cloned fragment in opposite orientation. Whole cell ELISA demonstrated that only when the orientation of transcription of the cloned fragment was identical with that of P1 promoter in the pBR322, could the CS3 antigen be expressed. And the test also demonstrated that the level of expression of CS3 antigen were different due to different on hosts. SDS-PAGE assay and Western blot identified that CS3 antigen had two different forms and the molecular weight were 15kD and 15.5 kD, respectively. Cloning of the gene encoding CS3 antigen permits us to study the regulation of its expression and to construct vaccine against ETEC.

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