System for in vitro transposition

Chemistry: molecular biology and microbiology – Process of mutation – cell fusion – or genetic modification – Introduction of a polynucleotide molecule into or...

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530350, 536 231, 536 241, C12N 1574, C07K 1300, C07H 2104

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059654430

ABSTRACT:
A system for in vitro transposition includes a donor DNA that includes a transposable element flanked by a pair of bacterial transposon Tn5 outside end repeat sequences, a target DNA into which the transposable element can transpose, and a modified Tn5 transposase having higher binding avidity to the outside end repeat sequences and being less likely to assume an inactive multimer form than wild type Tn5 transposase.

REFERENCES:
Benjamin, Howard W., "Excision of Tn10 from the donor site during transposition occurs by flush double-strand cleavages at the transposon termini," Proc. Natl. Acad. Sci. USA, 89:4648-4652 (May 1992).
Craigie, Robert, et al., "A defined system for the DNA strand-transfer reaction at the initiation of bacteriophage Mu transposition: Protein and DNA substrate requirements," Proc. Natl. Acad. Sci. USA, 82:7570-7574 (Nov. 1985).
de la Cruz, Norberto B., et al., "Characterization of the TN5 Transposase and Inhibitor Proteins: a Model for the Inhibition of Transposition," Journal of Bacterioilogy, 175(21):6932-6938 (Nov. 1993).
Jilk, Ross Alan, et al., "The Organization for the Outside End of Transposon Tn5," Journal of Bacteriology, 178(6):1671-1679 (Mar. 1996).
Lavoie, B.D., et al., "Transposition of Phage Mu DNA," Dept. of Biochemistry, Jniv. of Western Ontario. Current Topics in Microbiol and Immunol., vol. 204:83-102 (1995).
Mizuuchi, Kiyoshi, "In Vitro Transposition of Bacteriophage Mu: A Biochemical Approach to a Novel Replication Reaction," Cell, 35:785-794 (Dec. 1983).
Morisata, Donald, et al., "TN10 Transposition and Circle Formation In Vitro," Cell, 51:101-111 (Oct. 9, 1987).
Sakai, Janice, et al., "Identification and characterization of a pre-cleavage synaptic complex that is an early intermediate in TN10 transposition," The EMBO Journal, 14(17):4374-4383 (1995).
Weinreich, Michael D., "A Functional Analysis of the TN5 Transposase: Identification of Domains Required for DNA Binding and Multimerization," J. Mol. Biol., 241:166-177 (1994).
Weinreich, Michael D., et al., "Evidence that the cis preference of the Tn5 transposase is caused by nonproductive multimerization," Genes & Development, 8:2363-2374 (1994).
Wiegand, Torsten W., et al., "Characterization of Two Hypertransposing Tn5 Mutants," Journal of Bacteriology, 174(4):1229-1239 (Feb. 1992).
DeLong, A., et al., "Trans-Acting Transposase Mutant From Tn5," Proc. Natl. Acad. Sci. USA, 88:6072-6076 (1991).
Johnson, R.C., et al., "DNA Sequences at the Ends of Transposon Tn5 Required for Transposition," Nature 304:280-282 (1983).
Weigand, Torsten Walter, "Transposase Mutants That Increase the Transposition Frequency of Tn5 (Escherichia Coli)," Thesis abstract (1993).
Zhou, M., et al., "Three Types of Novel Mutations in the NH.sub.2 -Terminus of Tn5 Transposase: Structure/Function," Keystone Symposium abstract (1994), Supp 18B, p. 45 Journal of Cell Biology.

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