Identification of an EF-Tu protein that is periplasm-associated and processed in Neisseria gonorrhoeae

被引:23
作者
Porcella, SF
Belland, RJ
Judd, RC
机构
[1] NIAID, ROCKY MT LABS, NIH, HAMILTON, MT 59840 USA
[2] UNIV MONTANA, DIV BIOL SCI, MISSOULA, MT 59812 USA
来源
MICROBIOLOGY-SGM | 1996年 / 142卷
关键词
Neisseria gonorrhoeae; tufA; EF-Tu; processing;
D O I
10.1099/00221287-142-9-2481
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A 44 kDa protein is a dominant component of periplasmic extracts of Neisseria gonorrhoeae. Peptide sequence generated from a cyanogen-bromide-cleaved fragment of this protein indicated sequence homology with elongation factor-Tu (EF-Tu). Polyclonal antiserum was made against the 44 kDa protein purified from periplasm extracts of N. gonorrhoeae. The preabsorbed antiserum was immunoblotted against whole-cell lysates on two-dimensional gels. A 44 kDa protein and a smaller 37 kDa protein were recognized by this antiserum. A N. gonorrhoeae lambda phage DNA library was screened and a clone expressing a 44 kDa protein was identified. The DNA insert in this clone contained several genes homologous to genes contained in the str operon of Escherichia coli. One ORF product with a calculated molecular mass of 43 kDa was highly homologous to the EF-TuA of E. coli. A synthetic peptide antiserum specific for a portion of the C terminus of EF-Tu confirmed that the 37 kDa protein in whole-cell lysates of N. gonorrhoeae was a processed form of EF-Tu. Deletion of the tufA gene homologue in N. gonorrhoeae was attempted but was unsuccessful.
引用
收藏
页码:2481 / 2489
页数:9
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