Gene cloning using cDNA libraries in a differential competition hybridization strategy

Application to cloning XP-A related genes

A. Rinaldy, M. L. Dodson, T. L. Darling, Robert (Stephen) Lloyd

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

A competition hybridization strategy using size-cut cDNA libraries as both probe and competitor was designed for the cloning of genes whose mRNAs are either regulated transcriptionally or vary in abundance as a function of cell line or cell cycle. We used this strategy to construct cDNA libraries from a particular size fraction of mRNA from three members of a xeroderma pigmentosum (XP), complementation group A family. Size-cut cDNA libraries derived from the father's, mother's, and child's fibroblast cell line were used in a competition scheme to screen two λgt11 human cDNA libraries. Of the 15 positive λgt11 clones which have been characterized, at least 14 clones represent the same gene which is present in greater abundance in both the mother and father XP obligate heterozygotes relative to the homozygous affected child.

Original languageEnglish (US)
Pages (from-to)563-570
Number of pages8
JournalDNA
Volume7
Issue number8
StatePublished - 1988
Externally publishedYes

Fingerprint

Xeroderma Pigmentosum
Cloning
Gene Library
Organism Cloning
Genes
Cells
Fathers
Clone Cells
Mothers
Cell Line
Messenger RNA
Fibroblasts
Heterozygote
Cell Cycle

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Molecular Biology

Cite this

Gene cloning using cDNA libraries in a differential competition hybridization strategy : Application to cloning XP-A related genes. / Rinaldy, A.; Dodson, M. L.; Darling, T. L.; Lloyd, Robert (Stephen).

In: DNA, Vol. 7, No. 8, 1988, p. 563-570.

Research output: Contribution to journalArticle

@article{5c95ead32f0e4fb3aae3243f7d9e48c4,
title = "Gene cloning using cDNA libraries in a differential competition hybridization strategy: Application to cloning XP-A related genes",
abstract = "A competition hybridization strategy using size-cut cDNA libraries as both probe and competitor was designed for the cloning of genes whose mRNAs are either regulated transcriptionally or vary in abundance as a function of cell line or cell cycle. We used this strategy to construct cDNA libraries from a particular size fraction of mRNA from three members of a xeroderma pigmentosum (XP), complementation group A family. Size-cut cDNA libraries derived from the father's, mother's, and child's fibroblast cell line were used in a competition scheme to screen two λgt11 human cDNA libraries. Of the 15 positive λgt11 clones which have been characterized, at least 14 clones represent the same gene which is present in greater abundance in both the mother and father XP obligate heterozygotes relative to the homozygous affected child.",
author = "A. Rinaldy and Dodson, {M. L.} and Darling, {T. L.} and Lloyd, {Robert (Stephen)}",
year = "1988",
language = "English (US)",
volume = "7",
pages = "563--570",
journal = "DNA and Cell Biology",
issn = "1044-5498",
publisher = "Mary Ann Liebert Inc.",
number = "8",

}

TY - JOUR

T1 - Gene cloning using cDNA libraries in a differential competition hybridization strategy

T2 - Application to cloning XP-A related genes

AU - Rinaldy, A.

AU - Dodson, M. L.

AU - Darling, T. L.

AU - Lloyd, Robert (Stephen)

PY - 1988

Y1 - 1988

N2 - A competition hybridization strategy using size-cut cDNA libraries as both probe and competitor was designed for the cloning of genes whose mRNAs are either regulated transcriptionally or vary in abundance as a function of cell line or cell cycle. We used this strategy to construct cDNA libraries from a particular size fraction of mRNA from three members of a xeroderma pigmentosum (XP), complementation group A family. Size-cut cDNA libraries derived from the father's, mother's, and child's fibroblast cell line were used in a competition scheme to screen two λgt11 human cDNA libraries. Of the 15 positive λgt11 clones which have been characterized, at least 14 clones represent the same gene which is present in greater abundance in both the mother and father XP obligate heterozygotes relative to the homozygous affected child.

AB - A competition hybridization strategy using size-cut cDNA libraries as both probe and competitor was designed for the cloning of genes whose mRNAs are either regulated transcriptionally or vary in abundance as a function of cell line or cell cycle. We used this strategy to construct cDNA libraries from a particular size fraction of mRNA from three members of a xeroderma pigmentosum (XP), complementation group A family. Size-cut cDNA libraries derived from the father's, mother's, and child's fibroblast cell line were used in a competition scheme to screen two λgt11 human cDNA libraries. Of the 15 positive λgt11 clones which have been characterized, at least 14 clones represent the same gene which is present in greater abundance in both the mother and father XP obligate heterozygotes relative to the homozygous affected child.

UR - http://www.scopus.com/inward/record.url?scp=0023683294&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023683294&partnerID=8YFLogxK

M3 - Article

VL - 7

SP - 563

EP - 570

JO - DNA and Cell Biology

JF - DNA and Cell Biology

SN - 1044-5498

IS - 8

ER -