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[将粘性末端位点插入噬菌体M13的DNA及其在噬菌体λ蛋白质中的包装]

[Insertion of the cos-site into DNA of phage M13 and its packing in proteins of phage lambda].

作者信息

Zabarovskiĭ E R, Demirov D G, Nurbekov M K, Kiselev L L

出版信息

Bioorg Khim. 1986 Aug;12(8):1135-8.

PMID:3022756
Abstract

The cos-site of lambda phage from pHC79 cosmide is transferred to DNA from M13 mp18 phage. The recombinant DNA thus obtained (MC18) is efficiently packaged into lambda proteins in vitro. The BamHI-HindIII fragment of pGP588 (a pBR322 derivatives containing fragment of human DNA) is subcloned into MC18. Although this pGP588 fragment contains numerous Alu repeats, no essential rearrangements of the insert were revealed. The efficiency infection by recombinant DNA packaged with lambda proteins is about 1 X 10(5) pfu/microgram DNA, whereas in the similar conditions the efficiency of lambda EMBL3A was 1 X 10(6) pfu/microgram. It is assumed that the MC vectors might be suitable for cloning and sequencing large fragments either with cohesive or blunt ends. It opens also the way to construct genomic libraries in single-stranded phages.

摘要

来自黏粒载体pHC79的λ噬菌体黏性末端位点被转移到M13 mp18噬菌体的DNA上。由此获得的重组DNA(MC18)在体外能有效地被包装到λ蛋白中。pGP588(一种含有人类DNA片段的pBR322衍生物)的BamHI-HindIII片段被亚克隆到MC18中。尽管该pGP588片段含有大量Alu重复序列,但未发现插入片段有重要重排。用λ蛋白包装的重组DNA的感染效率约为1×10⁵噬菌斑形成单位/微克DNA,而在类似条件下λEMBL3A的效率为1×10⁶噬菌斑形成单位/微克。据推测,MC载体可能适用于克隆和测序具有黏性末端或平端的大片段。这也为在单链噬菌体中构建基因组文库开辟了道路。

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