Selective separation of virus proteins and double-stranded RNAs by SDS-KCl precipitation

Joseph K.K. Li; Todd Johnson; Yi Yuan Yang; Vicki Shore

doi:10.1016/0166-0934(89)90069-4

Selective separation of virus proteins and double-stranded RNAs by SDS-KCl precipitation

Joseph K.K. Li
, Todd Johnson
, Yi Yuan Yang
, Vicki Shore

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

20 Citations (Scopus)

Abstract

The total viral structural polypeptides and the double-stranded genomic RNAs of bluetongue virus can be selectively separated by a single SDS-KCl precipitation step. This simple, rapid and highly reproducible method enables greater than 95% recovery and purity of both viral proteins and dsRNAs within 30 min. The serotypic identity of the separated dsRNAs can be analyzed by SDS-PAGE electrophorogram immediately. After a single phenol/chloroform extraction, the dsRNA can also be used as hybridization probes, templates for molecular cloning and direct RNA sequencing. The SDS-KCl-precipitated viral proteins could be used readily for peptide mapping and as immunogens. Polyclonal and monoclonal antibodies raised against SDS-KCl-precipitated viral structural polypeptides were useful in Western immunoblots.

Original language	English
Pages (from-to)	3-15
Number of pages	13
Journal	Journal of Virological Methods
Volume	26
Issue number	1
DOIs	https://doi.org/10.1016/0166-0934(89)90069-4
Publication status	Published - Oct 1989
Externally published	Yes

Keywords

Protein isolation
SDS-KCl precipitation
dsRNA purification

ASJC Scopus subject areas

Virology

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10.1016/0166-0934(89)90069-4

Cite this

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title = "Selective separation of virus proteins and double-stranded RNAs by SDS-KCl precipitation",

abstract = "The total viral structural polypeptides and the double-stranded genomic RNAs of bluetongue virus can be selectively separated by a single SDS-KCl precipitation step. This simple, rapid and highly reproducible method enables greater than 95\% recovery and purity of both viral proteins and dsRNAs within 30 min. The serotypic identity of the separated dsRNAs can be analyzed by SDS-PAGE electrophorogram immediately. After a single phenol/chloroform extraction, the dsRNA can also be used as hybridization probes, templates for molecular cloning and direct RNA sequencing. The SDS-KCl-precipitated viral proteins could be used readily for peptide mapping and as immunogens. Polyclonal and monoclonal antibodies raised against SDS-KCl-precipitated viral structural polypeptides were useful in Western immunoblots.",

keywords = "Protein isolation, SDS-KCl precipitation, dsRNA purification",

author = "Li, \{Joseph K.K.\} and Todd Johnson and Yang, \{Yi Yuan\} and Vicki Shore",

note = "Funding Information: This study was supportedi n parts by Utah Agricultural Station Project 537 and by USDA grant 86-CRCR-1-2251.",

year = "1989",

month = oct,

doi = "10.1016/0166-0934(89)90069-4",

language = "English",

volume = "26",

pages = "3--15",

journal = "Journal of Virological Methods",

issn = "0166-0934",

publisher = "Elsevier B.V.",

number = "1",

}

TY - JOUR

T1 - Selective separation of virus proteins and double-stranded RNAs by SDS-KCl precipitation

AU - Li, Joseph K.K.

AU - Johnson, Todd

AU - Yang, Yi Yuan

AU - Shore, Vicki

N1 - Funding Information: This study was supportedi n parts by Utah Agricultural Station Project 537 and by USDA grant 86-CRCR-1-2251.

PY - 1989/10

Y1 - 1989/10

N2 - The total viral structural polypeptides and the double-stranded genomic RNAs of bluetongue virus can be selectively separated by a single SDS-KCl precipitation step. This simple, rapid and highly reproducible method enables greater than 95% recovery and purity of both viral proteins and dsRNAs within 30 min. The serotypic identity of the separated dsRNAs can be analyzed by SDS-PAGE electrophorogram immediately. After a single phenol/chloroform extraction, the dsRNA can also be used as hybridization probes, templates for molecular cloning and direct RNA sequencing. The SDS-KCl-precipitated viral proteins could be used readily for peptide mapping and as immunogens. Polyclonal and monoclonal antibodies raised against SDS-KCl-precipitated viral structural polypeptides were useful in Western immunoblots.

AB - The total viral structural polypeptides and the double-stranded genomic RNAs of bluetongue virus can be selectively separated by a single SDS-KCl precipitation step. This simple, rapid and highly reproducible method enables greater than 95% recovery and purity of both viral proteins and dsRNAs within 30 min. The serotypic identity of the separated dsRNAs can be analyzed by SDS-PAGE electrophorogram immediately. After a single phenol/chloroform extraction, the dsRNA can also be used as hybridization probes, templates for molecular cloning and direct RNA sequencing. The SDS-KCl-precipitated viral proteins could be used readily for peptide mapping and as immunogens. Polyclonal and monoclonal antibodies raised against SDS-KCl-precipitated viral structural polypeptides were useful in Western immunoblots.

KW - Protein isolation

KW - SDS-KCl precipitation

KW - dsRNA purification

UR - https://www.scopus.com/pages/publications/0024422973

UR - https://www.scopus.com/pages/publications/0024422973#tab=citedBy

U2 - 10.1016/0166-0934(89)90069-4

DO - 10.1016/0166-0934(89)90069-4

M3 - Article

C2 - 2556424

AN - SCOPUS:0024422973

SN - 0166-0934

VL - 26

SP - 3

EP - 15

JO - Journal of Virological Methods

JF - Journal of Virological Methods

IS - 1

ER -

Selective separation of virus proteins and double-stranded RNAs by SDS-KCl precipitation

Abstract

Keywords

ASJC Scopus subject areas

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