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将外源DNA体外包装到噬菌体T1的头部。

In vitro packaging of foreign DNA into heads of bacteriophage T1.

作者信息

Hug H, Hausmann R, Liebeschuetz J, Ritchie D A

出版信息

J Gen Virol. 1986 Feb;67 ( Pt 2):333-43. doi: 10.1099/0022-1317-67-2-333.

Abstract

The isolation of a collection of 44 morphologically T1-like phages is described. It is shown that these phages share some similarity with T1 in terms of cross-inactivation with anti-T1 serum, particle proteins and DNA packaging in vitro by the headful process. Virion DNA extracted from these phages was treated with T1 in vitro packaging extracts and the reaction mixtures were tested for the formation of infectious phage particles. The packaging efficiencies observed varied from about 1 to 100% of that of virion T1 DNA. Phage lambda virion DNA was packaged with an efficiency of between 0.01 and 2% (5 X 10(1) to 3 X 10(3) p.f.u./micrograms DNA), the shorter deleted derivative lambda L47 being packaged more efficiently than normal length lambda C1857 DNA. Virion DNA from phages T3 and T7 was also packaged at an efficiency similar to that for lambda. The in vitro packaging of T1 DNA requires the presence of the pac sequence which initiates headful packaging from a concatemeric precursor. The high efficiency of packaging DNA from some of the T1-like phages may indicate the presence of similar packaging sequences. However, in the case of lambda L47, which is known not to contain such a sequence, the in vitro DNA packaging reaction must occur by a secondary pathway unrelated to the headful mechanism.

摘要

本文描述了44种形态上类似T1的噬菌体的分离过程。结果表明,这些噬菌体在与抗T1血清的交叉失活、颗粒蛋白以及通过满头部包装过程进行体外DNA包装等方面与T1存在一些相似性。从这些噬菌体中提取的病毒粒子DNA用T1体外包装提取物进行处理,并检测反应混合物中感染性噬菌体颗粒的形成。观察到的包装效率为病毒粒子T1 DNA包装效率的约1%至100%。噬菌体λ病毒粒子DNA的包装效率在0.01%至2%之间(5×10¹至3×10³噬菌斑形成单位/微克DNA),较短的缺失衍生物λL47的包装效率高于正常长度的λC1857 DNA。噬菌体T3和T7的病毒粒子DNA的包装效率也与λ相似。T1 DNA的体外包装需要pac序列的存在,该序列从串联前体启动满头部包装。一些类似T1的噬菌体DNA的高效包装可能表明存在类似的包装序列。然而,就已知不包含此类序列的λL47而言,体外DNA包装反应必定通过与满头部机制无关的次要途径发生。

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