Cloning of cDNA encoding a 100 kDa nucleolar protein (nucleoline) of Chinese hamster ovary cells.
Nucleoline (100 kDa) is the major nucleolar protein in exponentially growing cells that behaves like a nucleolar organizer protein and plays a key role in rDNA transcription and prerRNA processing. We reported the isolation of 5 cDNA clones by probing a cDNA library, constructed in the expression vector lambda gt11, with a polyclonal serum raised against nucleoline. A new immunoassay, using hybrid proteins (beta gal-cDNA encoded protein) was developed to establish that the isolated cDNAs encoded parts of nucleoline. A further confirmation resulted from the sequence comparison between the cDNA encoded peptide and a 42 aa peptide isolated from rat nucleoline (1). The 5 cDNAs overlapped extensively and covered more than 90% of a full length cDNA. By probing a Northern blot with the 100 kDa cDNA, a 2650 nucleotide polyA+ RNA was detected that contained just enough information to code for nucleoline.
Lapeyre
B
B
Caizergues-Ferrer
M
M
Bouche
G
G
Amalric
F
F
D000595:Amino Acid Sequence; D000818:Animals; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000378:metabolism*; D004262:DNA Restriction Enzymes; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D008970:Molecular Weight; D009687:Nuclear Proteins; D000090243:Nucleophosmin; D010053:Ovary; D012261:Ribonucleoproteins / Q000235:genetics* / Q000302:isolation & purification; D001616:beta-Galactosidase / Q000235:genetics
D009687:Nuclear Proteins; D012261:Ribonucleoproteins; D000090243:Nucleophosmin; D004247:DNA; D004262:DNA Restriction Enzymes; D001616:beta-Galactosidase