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| Expression of fusion gene of tree Heat-stable Toxin I’ mutants and K99 adhesin of Enterotoxigenic Escherichia coli |
| GUO Le,LI Min,WANG Yu-jiong |
| Key Laboratory of Ministry of Education for Protection and Utilization of Special Biological Resource in Western China，School of Life Science，Ningxia University，Yinchuan 750021,China |
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Abstract Enterotoxigenic Escherichia coli(ETEC) is a major cause of diarrhea in animals and infants. The two most important virulence factors of ETEC are colonization factor antigens (CFAs), and the heat stable (ST) enterotoxin or heat labile (LT) enterotoxin.The recombinant plasmid pE3S(S)LK and pE3S(G)LK, containing three STI’s mutants and one colonization factor K99 gene were gotten successfully by PCR and double enzyme cleaving and inserting technology. The result of SDS-PAGE and Western blotting proved that the recombinant strain BL21(DE3)(pE3S(S)LK) and BL21(DE3)(pE3S(G)LK) could produce the fusion protein of 3STI(S)-K99 and 3STI(G)-K99 effectively which were specially recognized by anti-serum of enterotoxigenic Escherichia coli C83922.In addition, the recombinant protein of 3STΙ(S)-K99 and 3STΙ(G)-K99 tuned out to lose their biological toxicity of STΙ(The numerical value of G/C≤0.083) by the test of pouring stomach of sucking mice. It provides basic materials and theoretical introduction for prevent colibacillus diarrhea of newborn piglet and contributes to developing a new effective multivalent gene engneering vaccine.
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Received: 20 October 2009
Published: 29 April 2010
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