Loong Chan V, Louie H, Joe A
Department of Medical Genetics and Microbiology, and Department of Laboratory Medicine and Pathobiology, University of Toronto, 150 College Street, Toronto, Ontario,
Gene. 1998 Dec 28;225(1-2):131-41. doi: 10.1016/s0378-1119(98)00516-2.
Two Campylobacter jejuni genes with homology to the Escherichia coli flgF and flgG genes encoding two of the basal body rod proteins were isolated, and the nucleotide sequence was determined and analyzed. These two C. jejuni genes were shown, by Northern hybridization analysis, to function as a single operon (flgFG). Two transcriptional start sites were detected upstream of flgF, corresponding to the two RNA transcripts detected in the Northern blot. Western blot immunoassays using anti-FlgF and anti-FlgG antibodies demonstrated the synthesis of FlgF and FlgG proteins in C. jejuni and in Escherichia coli containing the C. jejuni flgF and flgG genes. Maxicell analysis and Western immunoblots using anti-FlgF antibodies to probe flgFG-encoded proteins in E. coli revealed the presence of a protein with a molecular mass of approximately the combined mass of the FlgF and FlgG proteins. Anti-FlgF antibodies detected in C. jejuni cell extracts the native FlgF protein and also a higher-molecular-weight protein that is likely encoded by the flgF and part of the flgG sequences.
分离出了空肠弯曲菌中两个与大肠杆菌flgF和flgG基因具有同源性的基因,这两个基因编码两种基体杆蛋白,并对其核苷酸序列进行了测定和分析。通过Northern杂交分析表明,这两个空肠弯曲菌基因作为一个单一操纵子(flgFG)发挥作用。在flgF上游检测到两个转录起始位点,与Northern印迹中检测到的两种RNA转录本相对应。使用抗FlgF和抗FlgG抗体的Western印迹免疫分析表明,在空肠弯曲菌以及含有空肠弯曲菌flgF和flgG基因的大肠杆菌中合成了FlgF和FlgG蛋白。使用抗FlgF抗体对大肠杆菌中flgFG编码的蛋白进行的Maxicell分析和Western免疫印迹显示,存在一种分子量约为FlgF和FlgG蛋白分子量之和的蛋白。抗FlgF抗体在空肠弯曲菌细胞提取物中检测到天然FlgF蛋白以及一种可能由flgF和部分flgG序列编码的高分子量蛋白。