A new bacteriophage P1-derived vector for the propagation of large human DNA fragments

We have designed a P1 vector (pCYPAC-1) for the introduction of recombinant DNA into E. coli using electroporation procedures. The new cloning system, P1-derived artificial chromosomes (PACs), was used to establish an initial 15,000 clone library with an average insert size of 130-150 kilobase pairs...

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Veröffentlicht in:	Nature genetics 1994-01, Vol.6 (1), p.84-89
Hauptverfasser:	loannou, Panayiotis A, Amemiya, Chris T, Garnes, Jeffrey, Kroisel, Peter M, Shizuya, Hiroaki, Chen, Chira, Batzer, Mark A, de Jong, Pieter J
Format:	Artikel
Sprache:	eng
Schlagworte:	Bacteriophage P1 - genetics Base Sequence Biological and medical sciences Biotechnology Cloning, Molecular DNA, Recombinant - genetics Escherichia coli Escherichia coli - genetics Fundamental and applied biological sciences. Psychology Gene Library Genetic engineering Genetic technics Genetic Vectors Humans In Situ Hybridization, Fluorescence Methods. Procedures. Technologies Molecular Sequence Data phage P1 Vectors (cloning, transfer, expression). Insertion sequences and transposons
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Zusammenfassung:	We have designed a P1 vector (pCYPAC-1) for the introduction of recombinant DNA into E. coli using electroporation procedures. The new cloning system, P1-derived artificial chromosomes (PACs), was used to establish an initial 15,000 clone library with an average insert size of 130-150 kilobase pairs (kb). No chimaerism has been observed in 34 clones, by fluorescence in situ hybridization. Similarly, no insert instability has been observed after extended culturing, for 20 clones. We conclude that the PAC cloning system will be useful in the mapping and detailed analysis of complex genomes.
ISSN:	1061-4036 1546-1718
DOI:	10.1038/ng0194-84