Patent ID: 11959069
Assignee: PROTEONIC BIOTECHNOLOGY IP B.V.
Field: Biotechnology (Chemistry)
Classification: CPC C | IPC C

Claim 15:
16. A method for enhancing transcription of a nucleotide sequence of interest in a eukaryotic cell, the method comprising the steps of:
i) providing a recombinant polynucleotide vector comprising the nucleotide surface of interest as an insert, and a vector backbone as defined in claim 1;
ii) amplifying the recombinant polynucleotide vector of step i) by transfecting the recombinant polynucleotide vector to a suitable microorganism and culturing said microorganism in a culture medium under conditions suitable for vector amplification to obtain amplified recombinant polynucleotide vector;
iii) isolating the amplified recombinant polynucleotide vector from the culture medium of step ii) to obtain isolated amplified recombinant polynucleotide vector;
iv) contacting the isolated amplified recombinant polynucleotide vector with cleavage means capable of cleaving the first plurality of cleavage sites to produce backbone fragments and the nucleotide surface of interest;
v) separating the nucleotide sequence of interest from the backbone fragments of step iv) to obtain an isolated nucleotide sequence of interest;
vi) integrating the isolated nucleotide sequence of interest into the genome of a eukaryotic cell to obtain a transgenic cell; and
vii) culturing the transgenic cell under conditions conducive to expression of the nucleotide sequence of interest.