DIFFERENTIATION OF MEDITERRANEAN PLUM POX VIRUS ISOLATES BY COAT PROTEIN-ANALYSIS
DIFFERENTIATION OF MEDITERRANEAN PLUM POX VIRUS ISOLATES BY COAT PROTEIN-ANALYSIS
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DOI：
10.1111/j.1365-3059.1994.tb00566.x
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发表时间：
1994-02-01
期刊：
影响因子：
2.7
通讯作者：
DIAZRUIZ, JR
中科院分区：
文献类型：
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作者：
LOPEZMOYA, JJ;CANTO, T;DIAZRUIZ, JR
采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）、肽图谱和Western印迹法，对来自不同地中海国家的6株李痘马铃薯Y病毒进行了比较。一个分离物（西班牙分离物3.3从李子）不同于其他人拥有一个较小的外壳蛋白（约34而不是36 kDa），在表面暴露的氨基末端区域可能缺失。使用局部病变宿主评估，蛋白水解后病毒的免疫力显著降低。建立蛋白酶消化条件以产生病毒外壳蛋白的28 kDa抗性核心。这种条件（较长的孵育时间或酶浓度的增加）不同于其他马铃薯Y病毒的温和报道。与病毒特异性单克隆抗体的生产和筛选的结果的影响进行了讨论。
Six isolates of plum pox potyvirus from different Mediterranean countries were compared by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), peptide mapping and Western blotting after improved purification of virions using a protease inhibitor cocktail that reduced coat protein degradation. One isolate (Spanish isolate 3.3 from plum) differed from the others in possessing a smaller coat protein (approximately 34 instead 36 kDa) with a possible deletion in the surface-exposed amino-terminal region. Infectivity of the viruses after proteolysis, assessed using a local lesion host, was significantly reduced. Protease digestion conditions were established to generate a 28 kDa resistant core of the viral coat protein. Such conditions (longer incubation times or an increase in the enzyme concentration) differed from the milder ones reported for other potyviruses. Implications of the results in relation to the production and screening of virus-specific monoclonal antibodies are discussed.