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
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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 -