| Nowsdays antimycotics has been wildly used, as a result, drug resistance becomes a problem which could not be ignored. One of the way to solve the problem is to search for new antimycotics. Endophytes are a potential source of novel chemistry and biology to assist in helping solve not only human health, but also plant and animal health problem, Now we can not take full adcantages of endophytes because many of their characters have not been understood, so reseaching on the anti-microbe endophyes not only brings vast social profits but also has realistic theoretic significance. An plant endophyte Paenibacillus kribbensis was isolated from the foliage of Aloe Chinenesis(Haw) Baker, whose secondary metabolite has been found to have a high inhlbitory on gram-postive (baeteria) such as staphylococcus aureus and bacillus subtilis. Our research group completed a comprehensive study of comparative genomics, decoding a greate number genome sequences of A11. Results showed that A11 contained gene ThiG,which chould code thiazole an antibacterial Substances.In order to reveal the relationship between candidate ThiG and Antibacterial activity of A11, the following experiments are conducted.1 Construction of a knockout vector of Paenibacillus kribbensis A11:the Kar gene cassette amplified from the pet28 a (+) vector was inserted to Bluescript SKâ…¡(+) to generate the PBSK-k vector. Then, The flanking DNA sequences to ThiG gene were amplified from the chromosomal DNA of endophyte A11 and were inserted to PBSK-k vector in turn, obtaining the knockout vector PBSK-lkr of endophyte A11 which was renamed as PBSK-ThiG.2 We use knockout vector PBSK-ThiG to transform competent cells plant of Paenibacillus kribbensis A11, and an ThiG gene deficient strains was get. The relation between antimicrobial activity of A11 and thiazole synthase through inhabition zone test .And the result reveals that thiazole synthase is not pivotal to antimicrobial activity of A11.
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