Bacterial strain for generating rhamnolipid biosurfactant and generated microbial inoculum thereof
A biological surface, rhamnolipid technology, applied in the field of biological high, can solve the problems of instability of plasmids, introduction of antibiotic markers, etc., to achieve the effect of protecting the ecological environment, promoting degradation and stable quality
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[0014] Primers were designed according to the rhamnolipid synthesis-related gene cluster (rhlABRI) sequence, and the forward primer was 5′-AGAT GCGGCCGC TTCGACACCGGAAAACC-3'; reverse primer is 5'-AGAT GCGGCCGC GGCATGGCGACTCCTC-3', a NotI restriction site was introduced at each end of the primer (the restriction sites are marked with underline). Using the genomic DNA of Pseudomonas aeruginosa BSFD5 as a template, the complete sequence of gene clusters related to rhamnolipid synthesis was amplified. After sequencing, it was registered in GenBank (accession number: HM190303). And ligated it into the vector pUTTns to construct pUTTns-rhlABRI, which was transformed into E.coli DH5α λpir E.coli DH5α λpir -pUTTns-rhlABRI. The rhlABRI gene cluster was integrated into the chromosome of Pseudomonas putida KT2440 by triparental conjugation, and the resistance gene was lost by screening the conjugants in LB medium supplemented with 10% (w / v) sucrose. The genetically engineered bact...
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