Finnzymes Oy

Other

Harri Savilahti

Anja Paatero

Mikko Frilander

Maria Pajunen

Xiaojuan Meng

Tuomas Tenkanen

Ville Tieaho

Timo Soininen

Kirsi Multanen

Hikka Turakainen

Tiina Rasila

Hilkka Turakainen

Meng Xiaojuan

The present invention provides a method for producing a hyperactive MuA transposase variant comprising at least one single-amino-acid change, the method comprising the steps of modifying the nucleic acid encoding wild type MuA transposase in at least one of the positions 59, 97, 160, 179, 233, 254, 258, 302, 335, 340, 345, 374, 447, 464, 478, 482, 483, 487, 495, 507, 539, 594 or 617 so that the modified nucleic acid encodes a MuA transposase variant comprising at least one single-amino-acid change in its amino acid sequence, wherein said single-amino-acid change results in higher enzyme activity of the variant when compared to the wild type MuA transposase. The present invention also provides hyperactive MuA transposases and kits comprising the same.

Method for producing hyperactive transposase mutants

The present invention relates to genetic engineering and especially to the use of DNA transposition complex of bacteriophage Mu. In particular, the invention provides a gene transfer system for eukaryotic cells, wherein in vitro assembled Mu transposition complexes are introduced into a target cell and subsequently transposition into a cellular nucleic acid occurs. The invention further provides a kit for producing insertional mutations into the genomes of eukaryotic cells. The kit can be used, e.g., to generate insertional mutant libraries.

Method for delivering nucleic acid into eukaryotic genomes

Delivery of nucleic acids into eukaryotic genomes using in vitro assembled Mu transposition complexes

The present invention describes an in vitro transposition-based methodology for generation of deletion derivatives of polypeptides. An artificial transposon containing at least partly within its transposon ends a modification with translation stop codons in three reading frames is provided. In the method, transposition complexes are assembled using the modified transposon and essentially random integrations into the target plasmid, containing a polypeptide coding nucleic acid of interest, are recovered as a plasmid pool. Subsequent manipulation steps including restriction enzyme digestions and ligation result in pools of mutant clones from which deletion derivatives of a polypeptide coding nucleic acid of interest and its respective deletion polypeptides could be produced.

Method and materials for producing deletion derivatives of polypeptides

The present invention relates to an in vitro method for providing unique templates for DNA sequencing and to a kit, which can be used, when employing the selection method of this invention. The examined DNA is subjected to a DNA transposition reaction and to an amplification reaction. The amplification reaction is carried out in the presence of a fixed primer and a selective primer having a complementary sequence to the end of a transposon DNA. A transposition reaction can be carried out also in the presence of a transposon or transposons, which have different ends enabling the design of selective primers for both ends. Each size of the amplification products represents one template for a sequencing reaction.

In vitro method for providing templates for DNA sequencing

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Helsinki, Finland

Harri Savilahti