Abstract：Stenotrophomonas maltophilia YHJ-1, isolated from the poultry factory, shows the high feather keratin degradation activity as well as many antibiotics resistances including kanamycin. In this study, a transposon plasmid pRH30 was developed using chloramphenicol gene to replace kanamycin gene with transposon plasmid pRL27 containing both hyperactive Tn5 transposase and kanamycin resistance gene and pMCm, carrying chloramphenicol cassette. Finally, an insertion library containing 1246 mutants was produced with pRH30 by biparental conjugation between donor E. coli UQ3201/pRH30 and recipient strain YHJ-1. The results of insertion mutants, subjected to detection of chloramphenicol resistance and PCR sequencing analysis, indicated that the library was reliable. The fact that 3 mutants were unable to degrade feather suggested that transposon mutagenesis with pRH30 was a highly efficient method for facilitating the isolation of genes and analyzing the molecular mechanism on feather-degrading of S. maltophili YHJ-1.
