TY - JOUR
T1 - Optimization of coliphage HK022 Integrase activity in human cells
AU - Malchin, Natalia
AU - Goltsman, Janna
AU - Dabool, Lital
AU - Gorovits, Rena
AU - Bao, Qiuye
AU - Dröge, Peter
AU - Yagil, Ezra
AU - Kolot, Mikhail
N1 - Funding Information:
Thanks to Dr. Orit Sagi-Asif for helping with the FACS analyses and to Michal Rosenzwaig for technical assistance. Research was supported by grant No. 2003394 to E.Y. and M.K from the United States–Israel Binational Science Foundation, Jerusalem, Israel.
PY - 2009/5/15
Y1 - 2009/5/15
N2 - The Integrase (Int) site-specific recombinase of coliphage HK022 catalyzes integrative and excisive DNA recombination between two attachment (att) sites in human cells without the need to supply the accessory proteins Integration Host Factor (IHF) and Excisionase (Xis). Previous work has shown that under these conditions, reactions in cis, i.e. both att sites are located on the same chromosome, can be detected without selection. However, recombination in trans, i.e. one att site positioned on a chromosome and the other on an episomal vector, was detected only after selection. Here we show that optimization of the int-HK022 gene for human codon usage according to the GeneOptimizer software algorithm, as well as addition of accessory proteins IHF and Xis improve the recombination efficiencies in human cells, such that recombinants in a trans reaction could be detected without selection.
AB - The Integrase (Int) site-specific recombinase of coliphage HK022 catalyzes integrative and excisive DNA recombination between two attachment (att) sites in human cells without the need to supply the accessory proteins Integration Host Factor (IHF) and Excisionase (Xis). Previous work has shown that under these conditions, reactions in cis, i.e. both att sites are located on the same chromosome, can be detected without selection. However, recombination in trans, i.e. one att site positioned on a chromosome and the other on an episomal vector, was detected only after selection. Here we show that optimization of the int-HK022 gene for human codon usage according to the GeneOptimizer software algorithm, as well as addition of accessory proteins IHF and Xis improve the recombination efficiencies in human cells, such that recombinants in a trans reaction could be detected without selection.
UR - http://www.scopus.com/inward/record.url?scp=63649102938&partnerID=8YFLogxK
U2 - 10.1016/j.gene.2009.02.013
DO - 10.1016/j.gene.2009.02.013
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AN - SCOPUS:63649102938
SN - 0378-1119
VL - 437
SP - 9
EP - 13
JO - Gene
JF - Gene
IS - 1-2
ER -