TY - JOUR
T1 - Structure of the dihydrofolate reductase gene in chinese hamster ovary cells
AU - Carothers, Adelaide M.
AU - Urlaub, Gail
AU - Ellis, Nathan
AU - Chasin, Lawrence A.
N1 - Funding Information:
ACKNOWLEDGEMENTS We thank Robert Schimke for providing pDHFR21 and Christian Simonsen for providing pDHFRll and unpublished mouse DNA sequence data. We are grateful for the valuable experimental assistance of Emmanuel Kas, Ted Widlanski, and Peter Karczmar. We thank Sally Lewing for preparation of this manuscript. This work was supported by NIH Grant No. GM34537 to L.A.C. and by a Damon Runyon - Walter Winchell Fellowship (DRG-446-F) to A.M.C.
PY - 1983/4/11
Y1 - 1983/4/11
N2 - 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.
AB - 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.
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U2 - 10.1093/nar/11.7.1997
DO - 10.1093/nar/11.7.1997
M3 - Article
C2 - 6300788
AN - SCOPUS:0021100734
SN - 0305-1048
VL - 11
SP - 1997
EP - 2012
JO - Nucleic acids research
JF - Nucleic acids research
IS - 7
ER -