Kieff E, Hoyer B, Bachenheimer S, Roizman B
J Virol. 1972 May;9(5):738-45. doi: 10.1128/JVI.9.5.738-745.1972.
The extent of homology between herpes simplex virus(1) and(2) (HSV-1 and HSV-2) deoxyribonucleic acid (DNA) was measured in two ways: (i) by determination of the relative rate of hybridization of labeled HSV-1 and HSV-2 DNA to excess unlabeled HSV-1 or HSV-2 DNA immobilized on filters and (ii) by determination of the rate of hybridization of labeled HSV-1 and HSV-2 DNA to excess unlabeled HSV-1 or HSV-2 DNA in solution. Approximately 40% of HSV-1 and HSV-2 DNA is homologous at hybridization temperatures 25 C below the melting temperature (T(m)) of HSV DNA (liquid-filter annealing). Lowering the temperature to 34 C below the T(m) increased the extent of homology to 46% (liquid annealing). The extent of base-pairing in HSV-1-HSV-2 heteroduplex DNA was determined by thermal chromatography on hydroxyapatite. Heteroduplexes of HSV-1 and HSV-2 DNA eluted in a single peak whose midpoint (Te(50)) was 10 C below that of the homoduplex. Conspicuously absent were heteroduplexes that eluted at more than 15 C below the Te(50) of the homoduplex. The data indicate the existence of a variable region of DNA (54%) with very little, if any, homology and an invariable region (46%) with relatively good (85%) matching of base pairs.
通过两种方法测定了单纯疱疹病毒1型和2型(HSV-1和HSV-2)脱氧核糖核酸(DNA)之间的同源程度:(i)通过测定标记的HSV-1和HSV-2 DNA与固定在滤膜上的过量未标记HSV-1或HSV-2 DNA的杂交相对速率;(ii)通过测定标记的HSV-1和HSV-2 DNA与溶液中过量未标记HSV-1或HSV-2 DNA的杂交速率。在比HSV DNA的解链温度(T(m))低25℃的杂交温度下,约40%的HSV-1和HSV-2 DNA是同源的(液相-滤膜退火)。将温度降至比T(m)低34℃时,同源程度增加到46%(液相退火)。通过在羟基磷灰石上进行热色谱分析来确定HSV-1-HSV-2异源双链DNA中的碱基配对程度。HSV-1和HSV-2 DNA的异源双链在一个单一峰中洗脱,其中点(Te(50))比同源双链的中点低10℃。明显不存在在比同源双链的Te(50)低超过15℃处洗脱的异源双链。数据表明存在一个DNA可变区(54%),其同源性极低(如果有的话),以及一个恒定区(46%),其碱基对匹配相对较好(85%)。