Patent ID: 11866724
Assignee: GENE BRIDGES GMBH
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 0:
1. A method for cloning an adenoviral sequence, wherein the adenoviral sequence is a full length adenoviral genome sequence, comprising:
a) providing a first linear nucleic acid molecule which comprises the full length adenoviral genome sequence;
b) providing a linearized medium copy plasmid which shares at least two regions of sequence homology with the first linear nucleic acid molecule; and
c) bringing the first linear nucleic acid molecule and the linearized medium copy plasmid into contact in the presence of a 5′ to 3′ exonuclease and an annealing protein such that the first linear nucleic acid molecule and the linearized medium copy plasmid recombine to form a circular plasmid containing the full length adenoviral genome sequence;
wherein the 5′ to 3′ exonuclease is full length RecE of SEQ ID NO:1412, or a protein with at least 95% sequence identity to SEQ ID NO:1412, and the annealing protein is RecT,
and wherein a first region of the at least two regions of sequence homology is a region of sequence homology with the 5′ ITR of the adenoviral sequence in the first linear nucleic acid molecule, and a second region of the at least two regions of sequence homology is a region of sequence homology with the 3′ ITR of the adenoviral sequence in the first linear nucleic acid molecule,
and wherein each of the first region and the second region is 40-80 nucleotides in length.