Bertani G
Jet Propulsion Laboratory, California Institute of Technology, Pasadena, California 91109, USA.
J Bacteriol. 1999 May;181(10):2992-3002. doi: 10.1128/JB.181.10.2992-3002.1999.
Strain PS of Methanococcus voltae (a methanogenic, anaerobic archaebacterium) was shown to generate spontaneously 4.4-kbp chromosomal DNA fragments that are fully protected from DNase and that, upon contact with a cell, transform it genetically. This activity, here called VTA (voltae transfer agent), affects all markers tested: three different auxotrophies (histidine, purine, and cobalamin) and resistance to BES (2-bromoethanesulfonate, an inhibitor of methanogenesis). VTA was most effectively prepared by culture filtration. This process disrupted a fraction of the M. voltae cells (which have only an S-layer covering their cytoplasmic membrane). VTA was rapidly inactivated upon storage. VTA particles were present in cultures at concentrations of approximately two per cell. Gene transfer activity varied from a minimum of 2 x 10(-5) (BES resistance) to a maximum of 10(-3) (histidine independence) per donor cell. Very little VTA was found free in culture supernatants. The phenomenon is functionally similar to generalized transduction, but there is no evidence, for the time being, of intrinsically viral (i.e., containing a complete viral genome) particles. Consideration of VTA DNA size makes the existence of such viral particles unlikely. If they exist, they must be relatively few in number;perhaps they differ from VTA particles in size and other properties and thus escaped detection. Digestion of VTA DNA with the AluI restriction enzyme suggests that it is a random sample of the bacterial DNA, except for a 0.9-kbp sequence which is amplified relative to the rest of the bacterial chromosome. A VTA-sized DNA fraction was demonstrated in a few other isolates of M. voltae.
沃氏甲烷球菌(一种产甲烷的厌氧古细菌)的菌株PS被证明能自发产生4.4千碱基对的染色体DNA片段,这些片段能完全抵抗脱氧核糖核酸酶,并且在与细胞接触时能对其进行基因转化。这种活性,在这里称为VTA(沃氏转移因子),影响所有测试的标记:三种不同的营养缺陷型(组氨酸、嘌呤和钴胺素)以及对BES(2-溴乙烷磺酸盐,一种产甲烷抑制剂)的抗性。VTA最有效的制备方法是通过培养过滤。这个过程破坏了一部分沃氏甲烷球菌细胞(其细胞质膜仅覆盖有一层S层)。VTA在储存时会迅速失活。VTA颗粒在培养物中的浓度约为每细胞两个。基因转移活性从每个供体细胞最低的2×10⁻⁵(对BES抗性)到最高的10⁻³(组氨酸非依赖性)不等。在培养上清液中发现的游离VTA很少。这种现象在功能上类似于广义转导,但目前没有证据表明存在内在病毒(即含有完整病毒基因组)颗粒。考虑到VTA DNA的大小,这种病毒颗粒不太可能存在。如果它们存在,数量肯定相对较少;也许它们在大小和其他特性上与VTA颗粒不同,因此未被检测到。用AluI限制性内切酶消化VTA DNA表明,它是细菌DNA的随机样本,除了一个相对于细菌染色体其他部分被扩增的0.9千碱基对序列。在沃氏甲烷球菌的其他一些分离株中也证明了存在VTA大小的DNA片段。