Screening of cDNA libraries with oligonucleotides as applied to signal transducing G proteins, receptors and effectors.
Screening of cDNA libraries constructed in phage or plasmids with oligonucleotide probes has become one of the preferred cloning techniques with the least number of false positive failures. In this article we present our current protocols for designing the procedure to detect cDNA inserts and isolate them. We illustrate with primary screens for G protein subunits and membrane receptors.
Abramowitz
J
J
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
Mattera
R
R
Liao
C F
CF
Olate
J
J
Perez-Ripoll
E
E
Birnbaumer
L
L
Codina
J
J
D010582:Bacteriophage lambda / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000032:analysis*; D004274:DNA, Recombinant / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D019204:GTP-Binding Proteins / Q000235:genetics; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000302:isolation & purification; D011956:Receptors, Cell Surface / Q000235:genetics
D004274:DNA, Recombinant; D009841:Oligonucleotides; D011956:Receptors, Cell Surface; D004247:DNA; D019204:GTP-Binding Proteins