Gene cloning using cDNA libraries in a differential competition hybridization strategy: application to cloning XP-A related genes.
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 lambda gt11 human cDNA libraries. Of the 15 positive lambda gt11 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.
Rinaldy
A
A
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville 37232.
Dodson
M L
ML
Darling
T L
TL
Lloyd
R S
RS
D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000032:analysis*; D006801:Humans; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis*
D012333:RNA, Messenger; D004247:DNA