Kitten T, Barbour A G
Department of Microbiology, University of Texas Health Science Center, San Antonio 78284.
Genetics. 1992 Oct;132(2):311-24. doi: 10.1093/genetics/132.2.311.
Borrelia hermsii, a spirochete which causes relapsing fever in humans and other mammals, eludes the immune response by antigenic variation of the "Vmp" proteins. This occurs by replacement of an expressed vmp gene with a copy of a silent vmp gene. Silent and expressed vmp genes are located on separate linear plasmids. To further characterize vmp recombination, copy numbers were determined for two linear plasmids and for the 1-megabase chromosome by comparing hybridization of probes to native DNA with hybridization to recombinant plasmids containing borrelial DNA. Plasmid copy numbers were also estimated by ethidium bromide fluorescence. Total cellular DNA content was determined by spectrophotometry. For borrelias grown in mice, copy numbers and 95% confidence intervals were 14 (12-17) for an expression plasmid, 8 (7-9) for a silent plasmid, and 16 (13-18) for the chromosome. Borrelias grown in broth medium had one-fourth to one-half this number of plasmids and chromosomes. Staining of cells with 4',6-diamidino-2-phenylindole revealed DNA to be distributed throughout most of the spirochete's length. These findings indicate that borrelias organize their total cellular DNA into several complete genomes and that cells undergoing serotype switches do one or more of the following: (1) coexpress Vmps from switched and unswitched expression plasmids for at least three to five generations, (2) suppress transcription from some expression plasmid copies, or (3) partition expression plasmids nonrandomly. The lower copy number of the silent plasmid indicates that nonreciprocal Vmp gene recombination may result from loss of recombinant silent plasmids by segregation.
赫氏疏螺旋体是一种可在人类和其他哺乳动物中引起回归热的螺旋体,它通过“Vmp”蛋白的抗原变异来逃避免疫反应。这是通过用一个沉默vmp基因的拷贝替换一个已表达的vmp基因来实现的。沉默和已表达的vmp基因位于不同的线性质粒上。为了进一步表征vmp重组,通过比较探针与天然DNA的杂交以及与含有疏螺旋体DNA的重组质粒的杂交,测定了两个线性质粒和1兆碱基染色体的拷贝数。还通过溴化乙锭荧光估计质粒拷贝数。通过分光光度法测定总细胞DNA含量。对于在小鼠体内生长的疏螺旋体,表达质粒的拷贝数及其95%置信区间为14(12 - 17),沉默质粒为8(7 - 9),染色体为16(13 - 18)。在肉汤培养基中生长的疏螺旋体,其质粒和染色体数量只有上述数量的四分之一到二分之一。用4',6-二脒基-2-苯基吲哚对细胞进行染色显示,DNA分布在螺旋体的大部分长度上。这些发现表明,疏螺旋体将其总细胞DNA组织成几个完整的基因组,并且经历血清型转换的细胞会进行以下一种或多种操作:(1) 从转换和未转换的表达质粒中共表达Vmp至少三到五代,(2) 抑制一些表达质粒拷贝的转录,或(3) 非随机分配表达质粒。沉默质粒较低的拷贝数表明,非相互性的Vmp基因重组可能是由于重组沉默质粒通过分离而丢失所致。