Isolation by polymerase chain reaction of a cDNA whose product partially complements the ultraviolet sensitivity of xeroderma pigmentosum group C cells

Tal Teitz; Michal Penner; Dalia Eli; Merav Stark; Mary Bakhanashvili; Tova Naiman; Dan Canaani

doi:10.1016/0378-1119(90)90316-J

Isolation by polymerase chain reaction of a cDNA whose product partially complements the ultraviolet sensitivity of xeroderma pigmentosum group C cells

Tal Teitz, Michal Penner, Dalia Eli, Merav Stark, Mary Bakhanashvili, Tova Naiman, Dan Canaani

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

17 Scopus citations

Abstract

A xeroderma pigmentosum (XP) cell line from complementation group C has been complemented to attain ultraviolet (UV) resistance and DNA repair proficiency, by transfection with a human expression cDNA library, followed by selection to UV resistance. We now show that the transfected cDNAs can be rescued from cellular DNA of a secondary transformant by its in vitro amplification using expression-vector-specific oligodeoxyribonucleotides as primers in a polymerase chain reaction. The amplified cDNAs were cloned into a mammalian expression vector. Their transfection into XP cells identified a single cDNA which specifically complemented the UV sensitivity of a group C-derived cell line to the same partial UV-resistance levels exhibited by the transformant from which the cDNAs were rescued.

Original language	English
Pages (from-to)	295-298
Number of pages	4
Journal	Gene
Volume	87
Issue number	2
DOIs	https://doi.org/10.1016/0378-1119(90)90316-J
State	Published - 15 Mar 1990
Externally published	Yes

Bibliographical note

Funding Information:
We thank H. Hofstetter for providing us with pCMVcat (the commercial use of the hCMV enhancer was patented by the Behringwerke on behalf of Dr. B. Fleckenstein and Dr. W. Schaffner), R. Palmiter for the gift of pMThGH, and D.L. Robberson, G.F. Saunders, W. Walker and J. Silva-Cudish for critical reading of the manuscript. We are grateful to the Laboratory of Endocrinology of Univer. sity Hospital 'Jose Eleuterio Gonz~lez' for the hGH radio= immunoassays. The work was supported in part by grants from 'Fondo de Estudios e Investigaci6n Ricardo J. Zevada', the Mexican Ministry of Public Education (SESIC) and the National Council of Science and Technology (CONACYT) ofthe Mexican Government. D.R.,P., D.E.R.-L., R.V.=M., and R.R.-S. thank the CONACYT for fellowships. We wish to express our gratitude to the School of Medicine of the Universidad A. de Nuevo Lebn for their continued support, and to M. P~ez and L. Cruz for the preparation of the manuscript.

Keywords

Recombinant DNA
expressible cDNA library
gene transfer
human disease
irradiation

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/0378-1119(90)90316-J

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title = "Isolation by polymerase chain reaction of a cDNA whose product partially complements the ultraviolet sensitivity of xeroderma pigmentosum group C cells",

abstract = "A xeroderma pigmentosum (XP) cell line from complementation group C has been complemented to attain ultraviolet (UV) resistance and DNA repair proficiency, by transfection with a human expression cDNA library, followed by selection to UV resistance. We now show that the transfected cDNAs can be rescued from cellular DNA of a secondary transformant by its in vitro amplification using expression-vector-specific oligodeoxyribonucleotides as primers in a polymerase chain reaction. The amplified cDNAs were cloned into a mammalian expression vector. Their transfection into XP cells identified a single cDNA which specifically complemented the UV sensitivity of a group C-derived cell line to the same partial UV-resistance levels exhibited by the transformant from which the cDNAs were rescued.",

keywords = "Recombinant DNA, expressible cDNA library, gene transfer, human disease, irradiation",

author = "Tal Teitz and Michal Penner and Dalia Eli and Merav Stark and Mary Bakhanashvili and Tova Naiman and Dan Canaani",

note = "Funding Information: We thank H. Hofstetter for providing us with pCMVcat (the commercial use of the hCMV enhancer was patented by the Behringwerke on behalf of Dr. B. Fleckenstein and Dr. W. Schaffner), R. Palmiter for the gift of pMThGH, and D.L. Robberson, G.F. Saunders, W. Walker and J. Silva-Cudish for critical reading of the manuscript. We are grateful to the Laboratory of Endocrinology of Univer. sity Hospital 'Jose Eleuterio Gonz~lez' for the hGH radio= immunoassays. The work was supported in part by grants from 'Fondo de Estudios e Investigaci6n Ricardo J. Zevada', the Mexican Ministry of Public Education (SESIC) and the National Council of Science and Technology (CONACYT) ofthe Mexican Government. D.R.,P., D.E.R.-L., R.V.=M., and R.R.-S. thank the CONACYT for fellowships. We wish to express our gratitude to the School of Medicine of the Universidad A. de Nuevo Lebn for their continued support, and to M. P~ez and L. Cruz for the preparation of the manuscript.",

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T1 - Isolation by polymerase chain reaction of a cDNA whose product partially complements the ultraviolet sensitivity of xeroderma pigmentosum group C cells

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AU - Penner, Michal

AU - Eli, Dalia

AU - Stark, Merav

AU - Bakhanashvili, Mary

AU - Naiman, Tova

AU - Canaani, Dan

N1 - Funding Information: We thank H. Hofstetter for providing us with pCMVcat (the commercial use of the hCMV enhancer was patented by the Behringwerke on behalf of Dr. B. Fleckenstein and Dr. W. Schaffner), R. Palmiter for the gift of pMThGH, and D.L. Robberson, G.F. Saunders, W. Walker and J. Silva-Cudish for critical reading of the manuscript. We are grateful to the Laboratory of Endocrinology of Univer. sity Hospital 'Jose Eleuterio Gonz~lez' for the hGH radio= immunoassays. The work was supported in part by grants from 'Fondo de Estudios e Investigaci6n Ricardo J. Zevada', the Mexican Ministry of Public Education (SESIC) and the National Council of Science and Technology (CONACYT) ofthe Mexican Government. D.R.,P., D.E.R.-L., R.V.=M., and R.R.-S. thank the CONACYT for fellowships. We wish to express our gratitude to the School of Medicine of the Universidad A. de Nuevo Lebn for their continued support, and to M. P~ez and L. Cruz for the preparation of the manuscript.

PY - 1990/3/15

Y1 - 1990/3/15

N2 - A xeroderma pigmentosum (XP) cell line from complementation group C has been complemented to attain ultraviolet (UV) resistance and DNA repair proficiency, by transfection with a human expression cDNA library, followed by selection to UV resistance. We now show that the transfected cDNAs can be rescued from cellular DNA of a secondary transformant by its in vitro amplification using expression-vector-specific oligodeoxyribonucleotides as primers in a polymerase chain reaction. The amplified cDNAs were cloned into a mammalian expression vector. Their transfection into XP cells identified a single cDNA which specifically complemented the UV sensitivity of a group C-derived cell line to the same partial UV-resistance levels exhibited by the transformant from which the cDNAs were rescued.

AB - A xeroderma pigmentosum (XP) cell line from complementation group C has been complemented to attain ultraviolet (UV) resistance and DNA repair proficiency, by transfection with a human expression cDNA library, followed by selection to UV resistance. We now show that the transfected cDNAs can be rescued from cellular DNA of a secondary transformant by its in vitro amplification using expression-vector-specific oligodeoxyribonucleotides as primers in a polymerase chain reaction. The amplified cDNAs were cloned into a mammalian expression vector. Their transfection into XP cells identified a single cDNA which specifically complemented the UV sensitivity of a group C-derived cell line to the same partial UV-resistance levels exhibited by the transformant from which the cDNAs were rescued.

KW - Recombinant DNA

KW - expressible cDNA library

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KW - human disease

KW - irradiation

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Isolation by polymerase chain reaction of a cDNA whose product partially complements the ultraviolet sensitivity of xeroderma pigmentosum group C cells

Abstract

Bibliographical note

Keywords

UN SDGs

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