Structure of the dihydrofolate reductase gene in chinese hamster ovary cells

Adelaide M. Carothers, Gail Urlaub, Nathan Ellis, Lawrence A. Chasin

Research output: Contribution to journalArticle

51 Scopus citations

Abstract

Overlapping recombinant λ1059 phages carrying regions of the dhfr locus from the amplified Chinese hamster ovary (CHO) cell clone MK42 have been isolated. In addition, dhfr cDNAs from this cell line have been cloned into plasmid pBR322. Restriction analysis of these recombinant molecules has led to a map of the Chinese hamster dhfr gene. This gene has a minimum size of 26 kb and contains six exons as defined by hybridization to a combination of mouse and CHO cDNA probes. The latter probes reveal 3' exonic sequences that are not present in mouse cDNA. The CHO dhfr gene thus extends about 700 bp further 3' than in the mouse, consistent with the larger size of the hamster mRNA. At least five intervening sequences are present, of approximate sizes: 0.3, 2.5, 8.6, 2.6 and 9.4 kb. Four sequences from highly repeated families are situated in introns within the dhfr gene. The overall structure of this gene is strikingly similar to that of the mouse. Evolutionary conservation of interrupted gene structure among mammals thus extends to genes that code for household enzymes as well as specialized or structural proteins.

Original languageEnglish (US)
Pages (from-to)1997-2012
Number of pages16
JournalNucleic acids research
Volume11
Issue number7
DOIs
StatePublished - Apr 11 1983
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

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