Stanton Thad B, Thompson M Greg, Humphrey Samuel B, Zuerner Richard L
Pre-Harvest Food Safety and Enteric Diseases Research Unit, National Animal Disease Center, Agricultural Research Service, United States Department of Agriculture, Ames, IA 50010, USA.
FEMS Microbiol Lett. 2003 Jul 29;224(2):225-9. doi: 10.1016/S0378-1097(03)00438-5.
VSH-1 is a mitomycin C-inducible, non-lytic, phage-like agent that packages random 7.5-kb fragments of the Brachyspira hyodysenteriae genome. VSH-1 is the first recognized mechanism for gene transfer between B. hyodysenteriae cells. To analyze the distribution of VSH-1 among spirochetes, a 344-bp probe for gene svp38, encoding the VSH-1 major head protein, was amplified by polymerase chain reaction and used in Southern blot hybridizations with genomic DNA from various spirochete genera. The svp38 probe hybridized to a 40-kb SalI-SmaI fragment of the B. hyodysenteriae B78(T) chromosome, indicating VSH-1 DNA insertion into the chromosome at a unique site. Restriction endonuclease digested DNAs of 27 spirochete strains representing six Brachyspira species (B. hyodysenteriae, B. innocens, B. pilosicoli, B. murdochii, B. intermedia, B. alvinipulli) contained a single fragment hybridizing with the svp38 probe. DNAs from spirochete species of the genera Treponema, Spirochaeta, Borrelia, and Leptospira did not hybridize with the probe. VSH-1-like agents appear to be widely distributed among Brachyspira species and, as has been demonstrated for B. hyodysenteriae, may serve as useful gene transfer agents for those other species.
VSH-1是一种丝裂霉素C诱导型、非裂解性、噬菌体样因子,它包裹着猪痢疾短螺旋体基因组的随机7.5kb片段。VSH-1是猪痢疾短螺旋体细胞间基因转移的首个被认可的机制。为了分析VSH-1在螺旋体中的分布情况,通过聚合酶链反应扩增了编码VSH-1主要头部蛋白的基因svp38的344bp探针,并将其用于与来自各种螺旋体属的基因组DNA进行Southern杂交。svp38探针与猪痢疾短螺旋体B78(T)染色体的一个40kb的SalI-SmaI片段杂交,表明VSH-1 DNA插入到染色体的一个独特位点。对代表六种短螺旋体物种(猪痢疾短螺旋体、无害短螺旋体、毛样短螺旋体、默多克短螺旋体、中间短螺旋体、阿尔文短螺旋体)的27个螺旋体菌株的限制性内切酶消化DNA进行分析,发现只有一个片段与svp38探针杂交。来自密螺旋体属、螺旋体属、疏螺旋体属和钩端螺旋体属的螺旋体物种的DNA与该探针不杂交。VSH-1样因子似乎在短螺旋体物种中广泛分布,并且正如在猪痢疾短螺旋体中所证明的那样,可能对其他物种也可作为有用的基因转移因子。