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 journalArticlepeer-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 languageEnglish
Pages (from-to)295-298
Number of pages4
JournalGene
Volume87
Issue number2
DOIs
StatePublished - 15 Mar 1990
Externally publishedYes

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

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