 Substrate for detection of mammalian 5-C-DNA methyltransferase |
| OF THE INVENTION "Oligomeric DNA" or ODN, as used herein, means a DNA having from 10 to 75 "purine ... |
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| Vinyl ethers of nucleoside analogs |
| OF THE INVENTION The compounds of the present invention can exist as optical isomers. Both the ... |
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| Nucleoside analogs |
| OF THE INVENTION The compounds of the present invention can exist as optical isomers. Both the ... |
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| Hydrophobic multicomponent heparin conjugates, a preparing method and a use thereof |
| To accomplish those objectives, the present invention provides hydrophobic muiticomponent heparin ... |
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| Gas permeable bioreactor and method of use |
| OF THE PREFERRED EMBODIMENTS Like numbered apparatus elements in FIGS. 1-11 are the same. R... |
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| Method for treating diseases of the inner ear using adamantane derivatives |
| OF THE INVENTION As already mentioned, the use of adamantane derivatives is possible or ... |
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| Polyalkylene oxide-containing urethane polyols with sulphonic acid groups |
| What is claimed is: 1. Sulphonic acid group-containing polyalkylene oxide-containing urethane ... |
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| Herpes treatment |
| I claim: 1. A method for the control of herpes virus infections in man which comprises contacting ... |
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| Dense star polymers and dendrimers |
| OF ILLUSTRATIVE EMBODIMENTS In the dense star polymers of the present invention, the core is ... |
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Vectors and methods for providing cells with additional nucleic acid material integrated in the genome of said cells
| Details |
Inventors: Plasterk, Ronald Hans Anton; Valerio, Domenico; Schouten, Govert Johan; van Luenen, Hendricus Gerhard Adrianus Maria; Vos, Jan C.;
Assignee: Het Nederlands Kanker Instituut (Amsterdam, NL); IntroGene B.V. (Leiden, NL)
Primary Examiner: Brusca; John S.
Assistant Examiner: Sandals; William
Attorney, Agent or Firm: Trask, Britt & Rossa
The present invention provides novel elements for improving genetic engineering techniques for producing recombinant nucleic acid molecules and/or recombinant cells. The novel elements are capable of integrating desired nucleic acid material into other nucleic acid materials, specifically into the genome of a host cell. The novel elements are derived from or based on transposons, in particular from the Tc/Mariner superfamily. In particular, the essential elements of Tc1 enabling excision and pasting of the desired nucleic acid material are provided, together with the relevant transposase activity in cis or in trans. |
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DETAILED DESCRIPTION OF THE INVENTION Tc1 belongs to the Tc1/mariner superfamily of transposons found in nematodes, arthropods and chordates (Henikoff 1992; Raddice et al.
1994; Robertson 1995; Plasterk 1995).
Both vertical and horizontal transfer have contributed to the spread of these elements throughout the animal kingdom (Robertson 1993; Radice et al.
1994; Robertson and Lampe 1995).
The widespread occurrence of the Tc1/mariner family of transposons can be taken as an indication for the absence of species-specific host factors which limit the transfer between different species.
Therefore, Tc1/mariner elements are attractive candidates for the development of gene delivery vectors.
Tc1-like elements are close to 1.
7 kb in length, have short inverted terminal repeats (ITR's) flanking a transposase gene and have the conserved sequence CAGT at their termini, flanked by TA representing the target site, which is duplicated upon integration (Van Luenen et al.
1994).
The element-encoded proteins share a homologous catalytic domain with bacterial transposases and retroviral integrases (Doak et al.
1994).
Tc1 from C.
elegans is a 1612 bp long transposon which has 54 bp inverted repeats flanking a gene encoding a 343 amino acid transposase (Enunons et al.
1983; Rosenzweig et al.
1983; Vos et al.
1993), that binds to the inverted repeats (Vos et al.
1993; Vos and Plasterk 1994).
The conserved hexanucleotide sequence, TACAGT, at the extreme termini of the element is not part of the transposase binding site, but is thought to play a role in catalysis of the transposition reaction (Vos and Plasterk 1994).
Here, we describe in vitro excision and transposition of Tc1 using an extract prepared from transgenic nematodes.
The minimal cis-requirements for transposition are defined and the target site choice in vitro is compared with that in vivo.
Furthermore, we demonstrate that recombinant transposase purified from E.
coli is capable of supporting transposition, showing that no other factors are essential for Tc1 transposition in vitro
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Gene Therapy 
