Patent ID: 8709778
Filing Date: 2014-04-29
Classification: C12N

Abstract:
1. A method of making a recombinant adenovirus vector, the method comprising the steps: (a) insertion of one or a group of endonuclease recognition sites into the genome of a helper adenovirus immediately downstream from adenovirus inverted terminal repeats; (b) amplification of the helper adenovirus, wherein the helper adenovirus contains adenovirus terminal protein bound to adenoviral DNA, wherein the adenoviral DNA is an inverted terminal repeat DNA sequence, wherein the adenovirus terminal protein bound to the inverted terminal repeat DNA sequence forms an adenovirus terminal protein-inverted terminal repeat DNA sequence complex; (c) purification of the adenovirus terminal protein-inverted terminal repeat DNA sequence complex from the helper adenovirus; and, (d) in vitro DNA ligation of the inverted terminal repeat DNA sequence of the adenovirus terminal protein-inverted terminal repeat DNA sequence complex to heterologous linear nucleic acid sequence by purified DNA ligase, wherein the heterologous linear nucleic acid sequence contains an adenovirus packaging signal, and wherein the adenovirus terminal protein-inverted terminal repeat DNA sequence complex ligated to said heterologous linear nucleic acid sequence is used to make recombinant adenovirus vector.