Open AccessCloning of cDNA encoding a 100 kDa nudeolar protein (nucleoline) of Chinese hamster ovary cells
Author(s) -
Bruno Lapeyre,
Michèle CaizerguesFerrer,
Gérard Bouche,
François Amalríc
Publication year - 1985
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/13.16.5805
Subject(s) - complementary dna , biology , microbiology and biotechnology , cdna library , rapid amplification of cdna ends , cloning (programming) , northern blot , chinese hamster ovary cell , nucleic acid sequence , molecular cloning , gene , genetics , cell culture , computer science , programming language
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.