CAJ | 학술논문

分别构建了两个烟草(Nicotiana tabacum L.)质体分裂基因NtFtsZ1和NtFtsZ2与编码绿色荧光蛋白的gfpS65A、V68L、S72A基因相融合的原核表达载体,并导入大肠杆菌( Escherichia coli ) JM109菌株中进行表达.全长NtFtsZs∶GFP融合蛋白在菌体中有规律地定位,暗示NtFtsZs能识别大肠杆菌潜在的分裂位点,并能与大肠杆菌的内源FtsZ发生聚合作用;融合蛋白的诱导表达抑制了宿主菌的分裂,形成了明显的丝状菌体,证明真核生物的 ftsZ 基因与大肠杆菌的 ftsZ 基因有相似的作用.同时构建了NtFtsZs不同缺失的原核表达载体,对这两个基因所编码蛋白不同结构域的功能做了初步分析.实验结果表明,烟草FtsZ蛋白的C端结构域与其在大肠杆菌细胞中的正确定位有关;而N端结构域与NtFtsZs∶GFP融合蛋白的聚合有关.
분별구건료량개연초(Nicotiana tabacum L.)질체분렬기인NtFtsZ1화NtFtsZ2여편마록색형광단백적gfpS65A、V68L、S72A기인상융합적원핵표체재체,병도입대장간균( Escherichia coli ) JM109균주중진행표체.전장NtFtsZs∶GFP융합단백재균체중유규률지정위,암시NtFtsZs능식별대장간균잠재적분렬위점,병능여대장간균적내원FtsZ발생취합작용;융합단백적유도표체억제료숙주균적분렬,형성료명현적사상균체,증명진핵생물적 ftsZ 기인여대장간균적 ftsZ 기인유상사적작용.동시구건료NtFtsZs불동결실적원핵표체재체,대저량개기인소편마단백불동결구역적공능주료초보분석.실험결과표명,연초FtsZ단백적C단결구역여기재대장간균세포중적정학정위유관;이N단결구역여NtFtsZs∶GFP융합단백적취합유관.
Two plastid division genes, NtFtsZ1 and NtFtsZ2 isolated from Nicotiana tabacum L. were fused with gfp and expressed in Escherichia coli filamentous bacteria indicated that the NtFtsZs could recognize the potential division sites in E. coli and be polymerized with heterogeneous division of host strain cells and resulted in the long filamentous bacterial morphology. These results suggested that eukaryotic ftsZs have similar function to their prokaryotic homologs. Meanwhile, the different deletions of motifs of NtFtsZs are also employed to investigate the functions of these proteins in E. coli . The results showed that the C-terminal domains of NtFtsZs were related to the correct localization of NtFtsZs in E. coli and the N-terminal domains of NtFtsZs were responsible for the polymerization of homogeneous and heterogeneous FtsZ proteins. The significance of these results in understanding the functions of NtFtsZs in plastid division were discussed.