Patrick Sheila, Parkhill Julian, McCoy Lisa J, Lennard Nicola, Larkin Michael J, Collins Martin, Sczaniecka Matylda, Blakely Garry
Microbiology and Immunobiology, The Queen's University of Belfast, Grosvenor Road, Belfast BT12 6BN, UK.
The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SA, UK.
Microbiology (Reading). 2003 Apr;149(Pt 4):915-924. doi: 10.1099/mic.0.26166-0.
The important opportunistic pathogen Bacteroides fragilis is a strictly anaerobic Gram-negative bacterium and a member of the normal resident human gastrointestinal microbiota. Our earlier studies indicated that there is considerable within-strain variation in polysaccharide expression, as detected by mAb labelling. Analysis of the genome sequence has revealed multiple invertible DNA regions, designated fragilis invertible (fin) regions, seven of which are upstream of polysaccharide biosynthesis loci and are approximately 226 bp in size. Using orientation-specific PCR primers and sequence analysis with populations enriched for one antigenic type, two of these invertible regions were assigned to heteropolymeric polysaccharides with different sizes of repeating units, as determined by PAGE pattern. The implication of these findings is that inversion of the fin regions switches biosynthesis of these polysaccharides off and on. The invertible regions are bound by inverted repeats of 30 or 32 bp with striking similarity to the Salmonella typhimurium H flagellar antigen inversion cross-over (hix) recombination sites of the invertible hin region. It has been demonstrated that a plasmid-encoded Hin invertase homologue (FinB), present in B. fragilis NCTC 9343, binds specifically to the invertible regions and the recombination sites have been designated as fragilis inversion cross-over (fix) sites.
重要的机会致病菌脆弱拟杆菌是一种严格厌氧的革兰氏阴性菌,也是人类正常胃肠道微生物群的成员。我们早期的研究表明,通过单克隆抗体标记检测到,菌株内多糖表达存在相当大的差异。基因组序列分析揭示了多个可反转DNA区域,称为脆弱拟杆菌可反转(fin)区域,其中七个位于多糖生物合成基因座的上游,大小约为226 bp。使用定向特异性PCR引物并对富集一种抗原类型的群体进行序列分析,通过PAGE图谱确定,其中两个可反转区域与具有不同重复单元大小的杂聚多糖相关。这些发现表明,fin区域的反转会开启和关闭这些多糖的生物合成。可反转区域由30或32 bp的反向重复序列结合,与鼠伤寒沙门氏菌H鞭毛抗原可反转hin区域的交叉(hix)重组位点具有惊人的相似性。已经证明,存在于脆弱拟杆菌NCTC 9343中的质粒编码Hin反转酶同源物(FinB)与可反转区域特异性结合,并且重组位点已被指定为脆弱拟杆菌交叉(fix)位点。