Patent ID: 6265159
Filing Date: 2001-07-24
Classification: C12N

Abstract:
A method for producing nested deletions in vitro in a desired DNA, comprising:1) preparing a vector containing a DNA fragment in which nested deletions are to he generated, and terminal repeats of a transposon;2) incubating said vector in vitro with a transposase and a DNA replication system, in a reaction solution containing 0.002 .mu.g/.mu.l to 0.04 .mu.g/.mu.l of said vector, producing a product;3) digesting the product of step 2) with a restriction enzyme which cleaves said vector containing the terminal repeats of the transposon and the contained DNA fragment only between one of the terminal repeats of the transposon and the contained DNA fragment;4) obtaining said vector containing said DNA fragment with nested deletions as a reaction product; and optionally,5) transforming a host cell with said reaction product and growing it; and6) recovering said reaction product which is the vector containing said DNA fragment with nested deletions from the grown host cell.