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Degradation of the viral strand of phiX174 parental replicative-form DNA in a rep- host.

机译:PHIX174父母复制形式DNA中的病毒链的降解。

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A progressive degradation of the parental viral strand label is observed upon infection of a Rep- mutant of Escherichia coli by 32P-labeled phiX174. Very little parental label remains in the RF (replicative form) by 47 min after infection. Concomitant with this degradation, replicative intermediates are formed which sediment at 21s, the rate of RF I (supercoiled-closed circular DNA), in a neutral sucrose gradient but which denature and sediment in alkaline gradients as single strands of unit size and larger. These denaturable 21s replicative intermediates have been shown previously to be RF molecules containing an elongated viral strand. Addition of chloramphenicol at 7 min after infection at 30 mug/ml, a concentration sufficient to block RF leads to SS (single strand) synthesis but not RF leads to RF synthesis in a wild-type host cell, reduced the amount of viral strand elongation but did not prevent viral strand degradation. The addition of chloramphenicol at 150 mug/ml at 7 min after infection totally prevents both the degradation of the parental label and the formation of the replicative intermediates with elongated tails. We infer that degradation of the viral strand requires the gene A-mediated nicking of the viral strand but not the concomitant elongation of the viral strand.
机译:通过32P标记的PHIX174感染大肠杆菌的牛突突变体,观察到父母病毒链标签的渐进降解。感染后47分钟,非常小的父母标签在RF(复制形式)中留在47分钟内。伴随该降解,形成重复的中间体,其在21s处,沉积在21s中,RF I(超填充圆形DNA)的速率在中性蔗糖梯度中,但在单位单位的单链和较大的单链中,碱性梯度的变性和沉积物。这些可变性21s复制中间体已被示出为含有细长病毒链的RF分子。在30麦克风/ mL感染后在7分钟内加入氯霉素,足以阻断RF导致SS(单链)合成的浓度,但不是RF导致野生型宿主细胞中的RF合成,减少了病毒链伸长的量但没有防止病毒链降解。在感染后,在7分钟内在150麦克霉/ ml下加入氯霉素,完全防止亲本标签的降解和具有细长尾部的复制中间体的形成。我们推断病毒链的降解需要基因a介导的病毒链切口,但不是病毒链的伴随伸长。

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  • 来源
    《Journal of Virology》 |1975年第4期|共6页
  • 作者

    K L Bowman; D S Ray;

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  • 收录信息 美国《科学引文索引》(SCI);
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