Cloning of terminal transferase cDNA by antibody screening.
A cDNA library was prepared from a terminal deoxynucleotidyltransferase-containing thymoma in the lambda phage vector lambda gt11. By screening plaques with anti-terminal transferase antibody, positive clones were identified of which some had beta-galactosidase-cDNA fusion proteins identifiable after electrophoretic fractionation by immunoblotting with anti-terminal transferase antibody. The predominant class of cross-hybridizing clones was determined to represent cDNA for terminal transferase by showing that one representative clone hybridized to a 2200-nucleotide mRNA in close-matched enzyme-positive but not to enzyme-negative cells and that the cDNA selected a mRNA that translated to give a protein of the size and antigenic characteristics of terminal transferase. Only a small amount of genomic DNA hybridized to the longest available clone, indicating that the sequence is virtually unique in the mouse genome.
D000818:Animals; D000906:Antibodies; D000936:Antigen-Antibody Complex; D010582:Bacteriophage lambda / Q000235:genetics; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism*; D004253:DNA Nucleotidylexotransferase / Q000032:analysis / Q000235:genetics*; D004254:DNA Nucleotidyltransferases / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors; D051379:Mice; D009693:Nucleic Acid Hybridization; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D013945:Thymoma / Q000201:enzymology*; D013953:Thymus Neoplasms / Q000201:enzymology; D001616:beta-Galactosidase / Q000235:genetics
D000906:Antibodies; D000936:Antigen-Antibody Complex; D012333:RNA, Messenger; D004247:DNA; D004254:DNA Nucleotidyltransferases; D004253:DNA Nucleotidylexotransferase; D001616:beta-Galactosidase