A simple purification of avian myeloblastosis virus reverse transcriptase for full-length transcription of 35 S RNA

Jeanne C. Myers; F. Ramirez; D. L. Kacian; M. Flood; S. Spiegelman

doi:10.1016/0003-2697(80)90044-5

A simple purification of avian myeloblastosis virus reverse transcriptase for full-length transcription of 35 S RNA

Jeanne C. Myers, F. Ramirez, D. L. Kacian, M. Flood, S. Spiegelman

Research output: Contribution to journal › Article › peer-review

6 Scopus citations

Abstract

Complete transcription of large RNA templates by avian myeloblastosis virus reverse transcriptase requires a purified and concentrated enzyme. This report describes a simple 2-day procedure consisting of a DEAE column, a carboxymethyl-Sepharose column, and a concentration step. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that the enzyme is free of contaminating protein and a series of rigorous assays reveal little if any exogenous ribonuclease or deoxyribonuclease activity. The reverse transcriptase purified by this method readily catalyzes synthesis of full-length complementary DNA from viral RNAs.

Original language	English
Pages (from-to)	88-96
Number of pages	9
Journal	Analytical Biochemistry
Volume	101
Issue number	1
DOIs	https://doi.org/10.1016/0003-2697(80)90044-5
State	Published - 1 Jan 1980
Externally published	Yes

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title = "A simple purification of avian myeloblastosis virus reverse transcriptase for full-length transcription of 35 S RNA",

abstract = "Complete transcription of large RNA templates by avian myeloblastosis virus reverse transcriptase requires a purified and concentrated enzyme. This report describes a simple 2-day procedure consisting of a DEAE column, a carboxymethyl-Sepharose column, and a concentration step. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that the enzyme is free of contaminating protein and a series of rigorous assays reveal little if any exogenous ribonuclease or deoxyribonuclease activity. The reverse transcriptase purified by this method readily catalyzes synthesis of full-length complementary DNA from viral RNAs.",

author = "Myers, {Jeanne C.} and F. Ramirez and Kacian, {D. L.} and M. Flood and S. Spiegelman",

note = "Funding Information: We express our appreciation to Dr. Joseph Beard and Dr. Dorothy Beard for generously supplying the avian myeloblastosis virus used in these studies. We thank Drs. T. Ohno, D. Mills, and C. Dobkin for many helpful suggestions and discussions and Ms. Susan LaFlamme for excellent technical assistance. This investigation was supported by Grant CA-02332 and Contract NOI-CP-1016 awarded by the National Cancer Institute.",

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AU - Kacian, D. L.

AU - Flood, M.

AU - Spiegelman, S.

N1 - Funding Information: We express our appreciation to Dr. Joseph Beard and Dr. Dorothy Beard for generously supplying the avian myeloblastosis virus used in these studies. We thank Drs. T. Ohno, D. Mills, and C. Dobkin for many helpful suggestions and discussions and Ms. Susan LaFlamme for excellent technical assistance. This investigation was supported by Grant CA-02332 and Contract NOI-CP-1016 awarded by the National Cancer Institute.

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N2 - Complete transcription of large RNA templates by avian myeloblastosis virus reverse transcriptase requires a purified and concentrated enzyme. This report describes a simple 2-day procedure consisting of a DEAE column, a carboxymethyl-Sepharose column, and a concentration step. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that the enzyme is free of contaminating protein and a series of rigorous assays reveal little if any exogenous ribonuclease or deoxyribonuclease activity. The reverse transcriptase purified by this method readily catalyzes synthesis of full-length complementary DNA from viral RNAs.

AB - Complete transcription of large RNA templates by avian myeloblastosis virus reverse transcriptase requires a purified and concentrated enzyme. This report describes a simple 2-day procedure consisting of a DEAE column, a carboxymethyl-Sepharose column, and a concentration step. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that the enzyme is free of contaminating protein and a series of rigorous assays reveal little if any exogenous ribonuclease or deoxyribonuclease activity. The reverse transcriptase purified by this method readily catalyzes synthesis of full-length complementary DNA from viral RNAs.

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A simple purification of avian myeloblastosis virus reverse transcriptase for full-length transcription of 35 S RNA

Abstract

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