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首页> 外文期刊>Journal of bacteriology >Physical and functional mapping of RP4-TOL plasmid recombinants: analysis of insertion and deletion mutants.
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Physical and functional mapping of RP4-TOL plasmid recombinants: analysis of insertion and deletion mutants.

机译：RP4-TOL质粒重组子的物理和功能定位：插入和缺失突变体的分析。

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Cleavage sites for the restriction endonucleases XhoI, BamHI, HindIII, and EcoRI were mapped on the pTN2 plasmid, a recombinant of TOL and RP4, which specifies the toluene-degrading enzymes in the same way as the wild-type TOL plasmid. The pTN2 plasmid, purified from a strain of Escherichia coli, contained the entire length of the RP4 plasmid (about 54 kilobase pairs [kb]) and the TOL segment (about 56 kb). The TOL segment is inserted at about 12 and 5 kb away from the EcoRI and BamHI cleavage sites of RP4, respectively. Cleavage sites for XhoI, BamHI, HindIII, and EcoRI were also mapped on an insertion mutant, pTN1, and two deletion mutants, pTN81 and pTN9. Analysis of pTN81 and pTN9 allowed estimation of the region of the gene cluster for benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase, as well as the region required for toluate oxygenase activity. Induction of TOL enzymes directed by pTN1 suggested the location and orientation of transcription of the gene cluster for catechol 2,3-oxygenase, 2-hydroxymuconic semialdehyde dehydrogenase, and 2-hydroxymuconic semialdehyde hydrolase. Analysis of strains carrying both pTN9 and a xylR mutant of the TOL plasmid demonstrated that xylR+ is trans dominant over xylR.
机译：限制性内切酶XhoI，BamHI，HindIII和EcoRI的切割位点被定位在pTN2质粒上，该质粒是TOL和RP4的重组体，它以与野生型TOL质粒相同的方式指定了降解甲苯的酶。从大肠杆菌菌株中纯化的pTN2质粒包含RP4质粒的全长（约54碱基对[kb]）和TOL片段（约56 kb）。将TOL片段分别插入距RP4的EcoRI和BamHI切割位点约12 kb和5 kb处。 XhoI，BamHI，HindIII和EcoRI的切割位点也定位在插入突变体pTN1和两个缺失突变体pTN81和pTN9上。通过对pTN81和pTN9的分析，可以估算出苄醇脱氢酶和苯甲醛脱氢酶的基因簇区域，以及甲苯氧化酶活性所需的区域。 pTN1指导的TOL酶的诱导表明邻苯二酚2,3-加氧酶，2-羟基粘康半醛脱氢酶和2-羟基粘康半醛水解酶的基因簇的转录位置和方向。对同时携带pTN9和TOL质粒的xylR突变体的菌株进行分析表明，xylR +比xylR具有反式优势。

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