CAJ | 학술논문

十字花科黑腐病菌8004菌株的XC3814基因与致病性和胞外多糖合成有关.文章将XC3814的启动子与报告基因sacB融合,构建了XC3814的表达报告质粒pL3814sac.将该质粒导入野生型菌株8004,获得了报告菌株8004/pL3814sac.利用转座子EZ::Tn5对报告菌株的基因组进行随机诱变,分离到3株耐蔗糖的突变体.分析发现其中的1株突变体是由EZ::Tn5插入到编号为XC3882的未知功能的基因所产生的.将由XC3814启动子与报告基因gusA融合得到的报告质粒pGUS3814分别导入8004菌株和XC3882的转座子Tn5gusA5插入突变体,测定比较pGUS3814的GUS表达水平,结果显示在XC3882突变体背景下GUS的表达水平比在野生型背景下降低81.3%,表明XC3814基因的表达水平受XC3882基因的影响.
십자화과흑부병균8004균주적XC3814기인여치병성화포외다당합성유관.문장장XC3814적계동자여보고기인sacB융합,구건료XC3814적표체보고질립pL3814sac.장해질립도입야생형균주8004,획득료보고균주8004/pL3814sac.이용전좌자EZ::Tn5대보고균주적기인조진행수궤유변,분리도3주내자당적돌변체.분석발현기중적1주돌변체시유EZ::Tn5삽입도편호위XC3882적미지공능적기인소산생적.장유XC3814계동자여보고기인gusA융합득도적보고질립pGUS3814분별도입8004균주화XC3882적전좌자Tn5gusA5삽입돌변체,측정비교pGUS3814적GUS표체수평,결과현시재XC3882돌변체배경하GUS적표체수평비재야생형배경하강저81.3%,표명XC3814기인적표체수평수XC3882기인적영향.
Xanthomonas campestris pv. campestris (Xcc) is the causal agent of the black rot disease of cruciferous plants. Our previous work had demonstrated that XC3814 is required for full virulence and extracellular polysaccharide production. In this work, the reporter plasmid pL3814sac was constructed by fusing the promoter region of XC3814 to the coding region of the gene sacB, and introduced into Xcc wild-type strain 8004. The resulted strain 8004/pL3814sac was mutagenized ran-domly by the transposon EZ::Tn5, and 3 mutant strains insensitive to sucrose were isolated. One of the mutants was due to the disruption of the open reading frame XC3882, which was assigned to code a hypothetical protein. To verify whether XC3882 has an impact on the expression level of XC3814, the reporter plasmid pGUS3814 was constructed by fusing the promoter region of XC3814 to the coding region of the gusA gene. This construct was introduced into the wild-type strain 8004 and the XC3882 mutant strain 190A10, which was derived from the transposon Tn5gusA5 insertion. The GUS activity, produced by pGUS3814 in the XC3882 mutant background, was reduced by 81.3% compared to that in the wild type back-ground. These results indicate that the expression of XC3814 is influenced by XC3882.