DNA sequencing vector with reversible insert

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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4352523, 43525233, 4353201, 935 29, 935 72, 935 73, C12Q 168, C12N 1500

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active

052272885

ABSTRACT:
A sequencing vector based on the M13 phage vector is disclosed which is particularly adapted for use in large scale DNA sequencing procedures. The vector includes a vector unique restriction site flanked by recognition sites, attP and attB, for the site specific recombination as catalyzed by a site specific recombination agent, lambda integrase. Using the vector, which incorporates the capability from M13 to replicate single stranded DNA for dideoxynucleotide sequencing, single stranded DNA of either strand may be selectively created since the orientation of any insert into the restriction site can readily be reversed by exposure of the vector to the recombination agent. This capability is particularly useful in large scale semi-random DNA sequencing in which the ability to selectively produce opposite strands is useful in filling gaps or ambiguities in large sequences.

REFERENCES:
patent: 4673640 (1987-06-01), Backman
Daniels et al., Lambda II, Weisburg et al., Ed., Appendix 2, pp. 519-576 (1983).
Hasan et al., Gene, 56:1, pp. 145-151 (1987) (Abstract only).
Hsu et al., Nature, 285 pp. 85-91 (1980).
Lee et al., J. Bacteriol., 172:3, pp. 1529-1538 (1990).
Mizuuchi et al., Nucleic Acids Research, 13:4, pp. 1193-1208 (1985).
Schroeder et al., Nucleic Acids Research, 10:1, pp. 69-84 (1982).
Yanisch-Perron et al., Gene, 33, pp. 103-119 (1985).

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