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Efficient expression of recombinant plasmids for colonization factor antigen 1 cfa 1 in escherichia coli



Efficient expression of recombinant plasmids for colonization factor antigen 1 cfa 1 in escherichia coli



Zhonghua Weishengwuxue He Mianyixue Zazhi 12(3): 157-161



We have constructed the plasmids pZLH42 and pZLH88 which contain the structural and regulatory genes for colonization factor antigen 1 (CFA/1). pZLH42 and pZLH88 have the same size but the inserting direction of the structural gene is just opposite. The expressing level of CFA/1 measured by ELISA was of much difference in different E. coli K12 strains harbouring plasmid pZLH42 or pZLH88. The expression of CFA/1 in E. coli RR1 or C600 was two-three times higher than E. coli H10407. The expression levels of E. coli Hb101 and E. coli H10407 were similar. The CFA/1 recombinant plasmids in E. coli C600 and HB101 were very stable when cultured in antibiotic-free medium. The plasmids in E. coli RR1 showed instability. After 100 generations when cultured in nonselective medium, 70% of cells lose the plasmids. The rabbit ileal loop test for LT toxin activity and suckling mice test for ST toxin activity were negative. So the CFA/1 recombinant clone could be a good live vaccine candidate for prevention of human diarrhea caused by ETEC bacteria.

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Accession: 007290917

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