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将来自质粒pKRP10、pKRP11和pKRP12的氯霉素、卡那霉素和四环素抗性基因分别插入质粒GFPmut2中gfp基因下游的PstI位点 ,得到gfp和不同抗性基因共存的重组质粒 ,转化Enterobactergergoviae 5 7- 7野生型菌株和耐铵工程菌E7后 ,得到既有抗生素抗性又在蓝光下呈现亮绿荧光的菌株 .用它们接种玉米后 ,利用这两种选择标记双重筛选重新分离到的细菌确定了接种菌在玉米幼苗根表面的定殖数目 ,同时用荧光显微镜观测了它们在玉米根表和土壤中的分布 .确证了GFP与抗生素抗性双标记定量环境中微生物的可靠性和GFP标记用于原位监测细菌分布的优越性 .图版 1图 1表 2参 15
The chloramphenicol, kanamycin and tetracycline resistance genes from plasmids pKRP10, pKRP11 and pKRP12 were respectively inserted into the PstI site downstream of the gfp gene in plasmid GFPmut2 to obtain recombinant plasmids in which gfp and different resistance genes coexist and transformed into Enterobacter germ oviae 5 7- 7 Wild-type strain and the ammonium-resistant engineering strain E7, both antibiotic resistance and bright green fluorescence under blue light were obtained.After inoculation with corn, these two selection markers were used to double screen for the bacteria The colonization of inoculated bacteria on the root surface of maize seedlings was confirmed, and their distribution in maize root surface and soil was observed by fluorescence microscopy.It was confirmed that the GFP and antibiotic resistant double-labeled quantitative microorganism reliability and GFP labeling Superiority for in-situ monitoring of bacterial distribution