USE OF DODECYL-BETA-D-MALTOSIDE IN THE PURIFICATION AND STABILIZATION OF RNA-POLYMERASE FROM BROME MOSAIC-VIRUS-INFECTED BARLEY

被引:57
作者
BUJARSKI, JJ [1 ]
HARDY, SF [1 ]
MILLER, WA [1 ]
HALL, TC [1 ]
机构
[1] UNIV WISCONSIN, DEPT HORT, MADISON, WI 53706 USA
关键词
D O I
10.1016/0042-6822(82)90105-2
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The activity and specificity of RNA-dependent RNA polymerase (replicase) isolated from brome mosaic virus-infected barley was enhanced by extraction with the nonionic detergent dodecyl-.beta.-D-maltoside. The enzyme was stable for at least 8 wk when stored at -70.degree. C. A further 100-fold purification was obtained by centrifugation through sucrose in the presence of detergent. The polymerase activity was associated with the pellet fraction; the template dependence and specificity were similar to those of the enzyme before sucrose purification. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed a 110 kilodalton protein in the purified pellet fraction from infected leaves that was absent from a similar fraction from healthy leaves. The protein had an identical electrophoretic mobility to that of protein 1a, the product of brome mosaic virus RNA 1 translation in vitro, and the profile of its tryptic polypeptides was very similar to that of protein 1a. RNA 1 apparently codes for the viral replicase, or a subunit thereof.
引用
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页码:465 / 473
页数:9
相关论文
共 17 条
[1]  
BOL JF, 1976, ANN INST PASTEUR MIC, VA127, P183
[2]   RNA-POLYMERASE FROM TOBACCO NECROSIS VIRUS-INFECTED AND UNINFECTED TOBACCO - PURIFICATION OF MEMBRANE-ASSOCIATED ENZYME [J].
FRAENKELCONRAT, H .
VIROLOGY, 1976, 72 (01) :23-32
[3]   NATURE OF PRODUCTS OF BROME MOSAIC-VIRUS RNA-POLYMERASE [J].
HADIDI, A .
VIROLOGY, 1974, 58 (02) :536-543
[4]   HIGHLY-ACTIVE TEMPLATE-SPECIFIC RNA-DEPENDENT RNA-POLYMERASE FROM BARLEY LEAVES INFECTED WITH BROME MOSAIC-VIRUS [J].
HARDY, SF ;
GERMAN, TL ;
LOESCHFRIES, LS ;
HALL, TC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1979, 76 (10) :4956-4960
[5]  
HELLENIUS A, 1979, METHOD ENZYMOL, V56, P734
[6]   RNA-DEPENDENT RNA-POLYMERASE OF TOBACCO PLANTS [J].
IKEGAMI, M ;
FRAENKELCONRAT, H .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1978, 75 (05) :2122-2124
[7]   CHANGES IN 3 OF THE 4 COAT PROTEINS OF ORAL POLIO VACCINE STRAIN DERIVED FROM TYPE-1 POLIOVIRUS [J].
KEW, OM ;
PALLANSCH, MA ;
OMILIANOWSKI, DR ;
RUECKERT, RR .
JOURNAL OF VIROLOGY, 1980, 33 (01) :256-263
[8]   VIRAL PROTEIN-SYNTHESIS IN BARLEY PROTOPLASTS INOCULATED WITH NATIVE AND FRACTIONATED BROME MOSAIC-VIRUS RNA [J].
KIBERSTIS, PA ;
LOESCHFRIES, LS ;
HALL, TC .
VIROLOGY, 1981, 112 (02) :804-808
[9]   PROPERTIES OF PRODUCTS SYNTHESIZED BY A DETERGENT-TREATED RNA-POLYMERASE PREPARATION FROM BARLEY LEAVES INFECTED WITH BROMEGRASS MOSAIC-VIRUS [J].
KUMMERT, J ;
SEMAL, J .
VIROLOGY, 1977, 77 (01) :212-220
[10]   CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].
LAEMMLI, UK .
NATURE, 1970, 227 (5259) :680-+