Characterization of the ftsZ cell division gene of Neisseria gonorrhoeae: expression in Escherichia coli and N. gonorrhoeae.
Characterization of the ftsZ cell division gene of Neisseria gonorrhoeae: expression in Escherichia coli and N. gonorrhoeae.
复制标题
淋病奈瑟氏菌 ftsZ 细胞分裂基因的表征：在大肠杆菌和淋病奈瑟氏菌中的表达。
DOI：
10.1007/s002030050002
复制
发表时间：
2000
影响因子：
2.8
通讯作者：
Dillon,JR
中科院分区：
文献类型：
--
作者：
Salimnia,H;Radia,A;Bernatchez,S;Beveridge,TJ;Dillon,JR
We cloned the cell division geneftsZof the gram-negative coccusNeisseria gonorrhoeae(Ng) strain CH811, characterized it genetically and phenotypically, and studied its localization inN. gonorrhoeaeandEscherichia coli(Ec). The 1,179-bp ORF offtsZNgencodes a protein with a predicted molecular mass of 41.5 kDa. Protein sequence alignments indicate that FtsZNgis similar to other FtsZ proteins and contains the conserved GTP binding motif. FtsZ homologues were identified in severalN. gonorrhoeaestrains and inNeisseria lactamica,Neisseria sicca,Neisseria polysaccharaeandNeisseria cinereaeither by Western blot or by PCR-Southern blot analysis. Attempts to inactivate theftsZNgon the chromosome failed, indicating that it is essential for gonococcal growth. FtsZNgwas synthesized in an in vitro transcription/translation system and was shown to be 43 kDa, the same size as in Western blots. Expression of theftsZNggene from nongonococcal promoters resulted in a filamentous phenotype inE. coli. Under controlled expression, the FtsZNg-GFP fusion protein localized at the mid-cell division site inE. coli.E. coliexpressing high levels of the FtsZNg-GFP fusion protein formed filaments and exhibited different fluorescent structures including helices, spiral tubules extending from pole to pole, and regularly spaced dots or bands that did not localize ¶at the middle of the cell. Expression of the FtsZNg-GFP fusion protein inN. gonorrhoeaeresulted in abnormal cell division as shown by electron microscopy. FtsZNg-GFP fusions were also expressed in a gonococcal background using a unique shuttle vector.
