The organization of the dihydrofolate reductase gene of baby hamster kidney fibroblasts.

نویسندگان

  • R G Schoner
  • J W Littlefield
چکیده

Using cloned DNA complementary to mouse dihydrofolate reductase (DHFR) mRNA, the organization of the hamster DHFR gene has been determined in two baby hamster kidney (BHK) cell lines, A5 and B1. A5 cells are highly methotrexate-resistant, containing 200-fold more copies of the DHFR gene than do the parental B1 cells. The DHFR gene has the same organization in A5 and B1 cells, suggesting that it has not been altered by the amplification process. The BHK DHFR gene spans a maximum of 10.7 kb and contains at least three introns. Thus the BHK DHFR gene is much smaller than the mouse DHFR gene, which has a minimum size of 42 kb and at least five introns. This striking size difference is probably due to much smaller introns in the BHK DHFR gene.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Genetic mutations in 57 and 58 codons gene of Plasmodium vivax dihydrofolate reductase

Introduction: The use of Sulfadoxine and pyrimethamine (SP) for treatment of vivax malaria is not common in most of malarious areas because of sensivity of this parasite to chloroquine. But, Plasmodium vivax isolates are exposed to SP because of mixed infection with P.falciparum and this subject has lead to emergence of mutations in P.vdhfr gene. As Plasmodium vivax is the most prevalent specie...

متن کامل

Molecular cloning of Chinese hamster dihydrofolate reductase-specific cDNA and the identification of multiple dihydrofolate reductase mRNAs in antifolate-resistant Chinese hamster lung fibroblasts.

ds cDNA from antifolate-resistant Chinese hamster lung fibroblast subline DC-3F/MQ19 was ligated to Eco RI and Sal I oligonucleotide linkers and cloned into Eco RI and Sal I digested pBR322. Transformed colonies containing dihydrofolate reductase (DHFR)-specific recombinant plasmid were identified by Grunstein Hogness assay using a Chinese hamster DHFR-specific cDNA probe. A recombinant plasmid...

متن کامل

Isolation and characterization of the multiple forms of dihydrofolate reductase from methotrexate-resistant hamster cells.

Dihydrofolate reductase, isolated from baby hamster kidney cells by affinity chromatography, gave multiple bands on analytical gel electrophoresis and electrofocusing. The number of bands could be reduced to two by forming a complex with the enzyme inhibitor, methotrexate. The two basic enzyme forms, called Form I and Form II, were obtained in large quantities by preparative disc electrophoresi...

متن کامل

Isolation of Chinese hamster cell mutants deficient in dihydrofolate reductase activity.

Mutants of Chinese hamster ovary cells lacking dihydrofolate reductase (tetrahydrofolate dehydrogenase, 7,8-dihydrofolate:NADP+ oxidoreductase; EC 1.5.1.3) activity were isolated after mutagenesis and exposure to high-specific-activity [3H]deoxyuridine as a selective agent. Fully deficient mutants could not be isolated starting with wild-type cells, but could readily be selected from a putative...

متن کامل

Isolation and characterization of a variant dihydrofolate reductase cDNA from methotrexate-resistant murine L5178Y cells.

Dihydrofolate reductase (DHFR) cDNA sequences were isolated from a methotrexate-resistant mouse L5178Y cell line previously shown to contain methotrexate-resistant dihydrofolate reductase enzyme activity. Specifically-primed reverse transcription products were amplified using the polymerase chain reaction and then cloned into a mammalian expression plasmid. Candidate clones were identified by r...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Nucleic acids research

دوره 9 23  شماره 

صفحات  -

تاریخ انتشار 1981