Isolating Cytochrome P450 cDNA and Genomic Clones

Library Screening with Synthetic DNA Oligomers

Christopher Hassett, Richard Ramsden, Curtis John Omiecinski

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

This chapter discusses the isolating cytochrome P450 complementary DNA (cDNA) and genomic clones. Cytochrome P450 enzymes are encoded by a large and complex superfamily of genes. Current P450 gene pools are believed to be the products of extensive duplication events, ultimately descended from one or more ancestral genes. Notable regions of conservation and divergence are apparent within P450 structures. Amino acid sequence conservation is found near a carboxy-terminal cysteine residue; this region is thought to function as the ligand to heme at the enzyme active site. Traditional methods of library screening fall within two categories: (1) antigenic detection using antibodies, and (2) homology to radiolabeled nucleic acid probes. Identification of target cDNAs by the interaction of antigens with antibody probe requires that the cloned DNA be inserted into an expression vector; the insert DNA must be in the correct orientation and reading frame.

Original languageEnglish (US)
Pages (from-to)291-301
Number of pages11
JournalMethods in enzymology
Volume206
Issue numberC
DOIs
StatePublished - Jan 1 1991

Fingerprint

Genomic Library
Oligomers
Cytochrome P-450 Enzyme System
Screening
Complementary DNA
Clone Cells
Genes
DNA
Conservation
Nucleic Acid Probes
Gene Pool
Reading Frames
Antibodies
Heme
Cysteine
Amino Acid Sequence
Catalytic Domain
Ligands
Antigens
Amino Acids

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Hassett, Christopher ; Ramsden, Richard ; Omiecinski, Curtis John. / Isolating Cytochrome P450 cDNA and Genomic Clones : Library Screening with Synthetic DNA Oligomers. In: Methods in enzymology. 1991 ; Vol. 206, No. C. pp. 291-301.
@article{9011b3b5722d49fdb7ce43881a82c315,
title = "Isolating Cytochrome P450 cDNA and Genomic Clones: Library Screening with Synthetic DNA Oligomers",
abstract = "This chapter discusses the isolating cytochrome P450 complementary DNA (cDNA) and genomic clones. Cytochrome P450 enzymes are encoded by a large and complex superfamily of genes. Current P450 gene pools are believed to be the products of extensive duplication events, ultimately descended from one or more ancestral genes. Notable regions of conservation and divergence are apparent within P450 structures. Amino acid sequence conservation is found near a carboxy-terminal cysteine residue; this region is thought to function as the ligand to heme at the enzyme active site. Traditional methods of library screening fall within two categories: (1) antigenic detection using antibodies, and (2) homology to radiolabeled nucleic acid probes. Identification of target cDNAs by the interaction of antigens with antibody probe requires that the cloned DNA be inserted into an expression vector; the insert DNA must be in the correct orientation and reading frame.",
author = "Christopher Hassett and Richard Ramsden and Omiecinski, {Curtis John}",
year = "1991",
month = "1",
day = "1",
doi = "10.1016/0076-6879(91)06099-O",
language = "English (US)",
volume = "206",
pages = "291--301",
journal = "Methods in Enzymology",
issn = "0076-6879",
publisher = "Academic Press Inc.",
number = "C",

}

Isolating Cytochrome P450 cDNA and Genomic Clones : Library Screening with Synthetic DNA Oligomers. / Hassett, Christopher; Ramsden, Richard; Omiecinski, Curtis John.

In: Methods in enzymology, Vol. 206, No. C, 01.01.1991, p. 291-301.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Isolating Cytochrome P450 cDNA and Genomic Clones

T2 - Library Screening with Synthetic DNA Oligomers

AU - Hassett, Christopher

AU - Ramsden, Richard

AU - Omiecinski, Curtis John

PY - 1991/1/1

Y1 - 1991/1/1

N2 - This chapter discusses the isolating cytochrome P450 complementary DNA (cDNA) and genomic clones. Cytochrome P450 enzymes are encoded by a large and complex superfamily of genes. Current P450 gene pools are believed to be the products of extensive duplication events, ultimately descended from one or more ancestral genes. Notable regions of conservation and divergence are apparent within P450 structures. Amino acid sequence conservation is found near a carboxy-terminal cysteine residue; this region is thought to function as the ligand to heme at the enzyme active site. Traditional methods of library screening fall within two categories: (1) antigenic detection using antibodies, and (2) homology to radiolabeled nucleic acid probes. Identification of target cDNAs by the interaction of antigens with antibody probe requires that the cloned DNA be inserted into an expression vector; the insert DNA must be in the correct orientation and reading frame.

AB - This chapter discusses the isolating cytochrome P450 complementary DNA (cDNA) and genomic clones. Cytochrome P450 enzymes are encoded by a large and complex superfamily of genes. Current P450 gene pools are believed to be the products of extensive duplication events, ultimately descended from one or more ancestral genes. Notable regions of conservation and divergence are apparent within P450 structures. Amino acid sequence conservation is found near a carboxy-terminal cysteine residue; this region is thought to function as the ligand to heme at the enzyme active site. Traditional methods of library screening fall within two categories: (1) antigenic detection using antibodies, and (2) homology to radiolabeled nucleic acid probes. Identification of target cDNAs by the interaction of antigens with antibody probe requires that the cloned DNA be inserted into an expression vector; the insert DNA must be in the correct orientation and reading frame.

UR - http://www.scopus.com/inward/record.url?scp=0026319864&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026319864&partnerID=8YFLogxK

U2 - 10.1016/0076-6879(91)06099-O

DO - 10.1016/0076-6879(91)06099-O

M3 - Article

VL - 206

SP - 291

EP - 301

JO - Methods in Enzymology

JF - Methods in Enzymology

SN - 0076-6879

IS - C

ER -