ISSN:
1432-072X
Schlagwort(e):
Key words Bacteria
;
Cloning
;
FtsZ
;
Cell division
;
¶Neisseria gonorrhoeae
;
Localization
;
Expression
;
Green fluorescent protein
Quelle:
Springer Online Journal Archives 1860-2000
Thema:
Biologie
Notizen:
Abstract We cloned the cell division gene ftsZ of the gram-negative coccus Neisseria gonorrhoeae (Ng) strain CH811, characterized it genetically and phenotypically, and studied its localization in N. gonorrhoeae and Escherichia coli (Ec). The 1,179-bp ORF of ftsZ Ng encodes a protein with a predicted molecular mass of 41.5 kDa. Protein sequence alignments indicate that FtsZNg is similar to other FtsZ proteins and contains the conserved GTP binding motif. FtsZ homologues were identified in several N. gonorrhoeae strains and in Neisseria lactamica, Neisseria sicca, Neisseria polysaccharae and Neisseria cinerea either by Western blot or by PCR-Southern blot analysis. Attempts to inactivate the ftsZ Ng on the chromosome failed, indicating that it is essential for gonococcal growth. FtsZNg was synthesized in an in vitro transcription/translation system and was shown to be 43 kDa, the same size as in Western blots. Expression of the ftsZ Ng gene from nongonococcal promoters resulted in a filamentous phenotype in E. coli. Under controlled expression, the FtsZNg-GFP fusion protein localized at the mid-cell division site in E. coli. E. coli expressing 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 in N. gonorrhoeae resulted in abnormal cell division as shown by electron microscopy. FtsZNg-GFP fusions were also expressed in a gonococcal background using a unique shuttle vector.
Materialart:
Digitale Medien
URL:
http://dx.doi.org/10.1007/s002030050002
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