Patent ID: 9309518
Filing Date: 2016-04-12
CPC Classification: C12N

Claim Text:
1. A method for sub-cloning and stabilizing a nucleic acid sequence of interest into a cell this method comprising the steps of: a) preparing DNA of a receiving vector comprising: b) performing recombination between the said receiving vector and a donor vector, said donor vector comprising the nucleic acid sequence of interest being located between a second pair of two different recombination or restriction sites or recombination arms wherein, this second pair of recombination or restriction sites or recombination arms being compatible with the first pair of two recombination sites or recombination arms of the receiving vector; c) obtaining a chimeric nucleic acid molecule comprising the nucleic acid sequence of interest; d) transforming the obtained recombination mixture in cell(s) carrying a nucleotide sequence encoding the second poison protein; e) obtaining a synthesis of the first poison protein and second poison protein and of the antidote protein to the said second poison protein in the cells; f) selecting cells surviving to the toxic activity of the first poison protein and the second poison protein; and g) growing and collecting recovered surviving cells comprising the nucleic acid sequence of interest, under selective pressure of the second poison protein.