Optimization of coliphage HK022 Integrase activity in human cells

Natalia Malchin; Janna Goltsman; Lital Dabool; Rena Gorovits; Qiuye Bao; Peter Dröge; Ezra Yagil; Mikhail Kolot

doi:10.1016/j.gene.2009.02.013

Optimization of coliphage HK022 Integrase activity in human cells

Natalia Malchin, Janna Goltsman, Lital Dabool, Rena Gorovits, Qiuye Bao, Peter Dröge, Ezra Yagil, Mikhail Kolot^*

^*Corresponding author for this work

Institute of Plant Sciences and Genetics in Agriculture

Research output: Contribution to journal › Article › peer-review

8 Scopus citations

Abstract

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.

Original language	English
Pages (from-to)	9-13
Number of pages	5
Journal	Gene
Volume	437
Issue number	1-2
DOIs	https://doi.org/10.1016/j.gene.2009.02.013
State	Published - 15 May 2009

Bibliographical note

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.

Access to Document

10.1016/j.gene.2009.02.013

Cite this

@article{7cb09dfc3ab748a0a32c0de1442f522e,

title = "Optimization of coliphage HK022 Integrase activity in human cells",

abstract = "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.",

author = "Natalia Malchin and Janna Goltsman and Lital Dabool and Rena Gorovits and Qiuye Bao and Peter Dr{\"o}ge and Ezra Yagil and Mikhail Kolot",

note = "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.",

year = "2009",

month = may,

day = "15",

doi = "10.1016/j.gene.2009.02.013",

language = "אנגלית",

volume = "437",

pages = "9--13",

journal = "Gene",

issn = "0378-1119",

publisher = "Elsevier",

number = "1-2",

}

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

M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???

C2 - 19268511

AN - SCOPUS:63649102938

SN - 0378-1119

VL - 437

SP - 9

EP - 13

JO - Gene

JF - Gene

IS - 1-2

ER -

Optimization of coliphage HK022 Integrase activity in human cells

Abstract

Bibliographical note

Access to Document

Other files and links

Fingerprint

Cite this