Isolation and expression of the Pneumocystis carinii dihydrofolate reductase gene

J C Edman; U Edman; Mi-Mi Cao; B Lundgren; J A Kovacs; D V Santi

Isolation and expression of the Pneumocystis carinii dihydrofolate reductase gene

J C Edman, U Edman, Mi-Mi Cao, B Lundgren, J A Kovacs, D V Santi

144 Citationer (Scopus)

Abstract

Pneumocystis carinii dihydrofolate reductase (DHFR; 5,6,7,8-tetrahydrofolate: NADP+ oxidoreductase, EC 1.5.1.3) cDNA sequences have been isolated by their ability to confer trimethoprim resistance to Escherichia coli. Consistent with the recent conclusion that P. carinii is a member of the Fungi, sequence analysis and chromosomal localization show that DHFR is neither physically nor genetically linked to thymidylate synthase. Expression of recombinant P. carinii DHFR in heterologous hosts provides an abundant source of the enzyme that may form a basis for the development of new therapies for this enigmatic pathogen. Studies with the recombinant enzyme show that trimethoprim is a very poor inhibitor of P. carinii DHFR and, in fact, is a more potent inhibitor of human DHFR.

Originalsprog	Engelsk
Tidsskrift	Proceedings of the National Academy of Sciences of the United States of America
Vol/bind	86
Udgave nummer	22
Sider (fra-til)	8625-9
Antal sider	5
ISSN	0027-8424
Status	Udgivet - nov. 1989

Citationsformater

@article{c7f82314b4bc44378993d4e84aa8df2f,

title = "Isolation and expression of the Pneumocystis carinii dihydrofolate reductase gene",

abstract = "Pneumocystis carinii dihydrofolate reductase (DHFR; 5,6,7,8-tetrahydrofolate: NADP+ oxidoreductase, EC 1.5.1.3) cDNA sequences have been isolated by their ability to confer trimethoprim resistance to Escherichia coli. Consistent with the recent conclusion that P. carinii is a member of the Fungi, sequence analysis and chromosomal localization show that DHFR is neither physically nor genetically linked to thymidylate synthase. Expression of recombinant P. carinii DHFR in heterologous hosts provides an abundant source of the enzyme that may form a basis for the development of new therapies for this enigmatic pathogen. Studies with the recombinant enzyme show that trimethoprim is a very poor inhibitor of P. carinii DHFR and, in fact, is a more potent inhibitor of human DHFR.",

keywords = "Amino Acid Sequence, Base Sequence, Cloning, Molecular, DNA, Bacterial, Escherichia coli, Gene Expression, Gene Library, Genes, Bacterial, Humans, Molecular Sequence Data, Oligonucleotide Probes, Plasmids, Pneumocystis, Sequence Homology, Nucleic Acid, Tetrahydrofolate Dehydrogenase",

author = "Edman, {J C} and U Edman and Mi-Mi Cao and B Lundgren and Kovacs, {J A} and Santi, {D V}",

year = "1989",

month = nov,

language = "English",

volume = "86",

pages = "8625--9",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

issn = "0027-8424",

publisher = "National Academy of Sciences",

number = "22",

}

TY - JOUR

T1 - Isolation and expression of the Pneumocystis carinii dihydrofolate reductase gene

AU - Edman, J C

AU - Edman, U

AU - Cao, Mi-Mi

AU - Lundgren, B

AU - Kovacs, J A

AU - Santi, D V

PY - 1989/11

Y1 - 1989/11

N2 - Pneumocystis carinii dihydrofolate reductase (DHFR; 5,6,7,8-tetrahydrofolate: NADP+ oxidoreductase, EC 1.5.1.3) cDNA sequences have been isolated by their ability to confer trimethoprim resistance to Escherichia coli. Consistent with the recent conclusion that P. carinii is a member of the Fungi, sequence analysis and chromosomal localization show that DHFR is neither physically nor genetically linked to thymidylate synthase. Expression of recombinant P. carinii DHFR in heterologous hosts provides an abundant source of the enzyme that may form a basis for the development of new therapies for this enigmatic pathogen. Studies with the recombinant enzyme show that trimethoprim is a very poor inhibitor of P. carinii DHFR and, in fact, is a more potent inhibitor of human DHFR.

AB - Pneumocystis carinii dihydrofolate reductase (DHFR; 5,6,7,8-tetrahydrofolate: NADP+ oxidoreductase, EC 1.5.1.3) cDNA sequences have been isolated by their ability to confer trimethoprim resistance to Escherichia coli. Consistent with the recent conclusion that P. carinii is a member of the Fungi, sequence analysis and chromosomal localization show that DHFR is neither physically nor genetically linked to thymidylate synthase. Expression of recombinant P. carinii DHFR in heterologous hosts provides an abundant source of the enzyme that may form a basis for the development of new therapies for this enigmatic pathogen. Studies with the recombinant enzyme show that trimethoprim is a very poor inhibitor of P. carinii DHFR and, in fact, is a more potent inhibitor of human DHFR.

KW - Amino Acid Sequence

KW - Base Sequence

KW - Cloning, Molecular

KW - DNA, Bacterial

KW - Escherichia coli

KW - Gene Expression

KW - Gene Library

KW - Genes, Bacterial

KW - Humans

KW - Molecular Sequence Data

KW - Oligonucleotide Probes

KW - Plasmids

KW - Pneumocystis

KW - Sequence Homology, Nucleic Acid

KW - Tetrahydrofolate Dehydrogenase

M3 - Journal article

C2 - 2682653

SN - 0027-8424

VL - 86

SP - 8625

EP - 8629

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 22

ER -

Isolation and expression of the Pneumocystis carinii dihydrofolate reductase gene

Abstract

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