Ross Jerri, Ostlund Eileen N, Cao Dianjun, Tatsumi Masatoshi, Hoshino Yasutaka
Epidemiology Section, Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-8026, USA.
J Clin Virol. 2008 Aug;42(4):374-80. doi: 10.1016/j.jcv.2008.03.019. Epub 2008 May 23.
During the course of development and characterization of various rotavirus reassortants, we found that the relative position of the gene encoding neutralization and protective antigen VP7 of certain rotavirus strains in a PAGE gel was influenced by the concentration of acrylamide.
We investigated systematically various factors that affected the relative position of the rotavirus VP7 gene in a PAGE gel.
We analyzed dsRNAs of selected rotavirus strains bearing G1, G2, G3 or G9 specificity by PAGE at varying concentrations of acrylamide.
We demonstrated that the relative position of the VP7 gene of three G2 strains varied depending upon the concentration of acrylamide in a PAGE gel, which occurred not only in a homologous G2 virus gene background but also in a heterologous G3 virus gene background; and the VP7 gene bearing G1, G3, G4 or G9 specificity did not display this phenomenon when the PAGE running conditions were varied.
The concentration of acrylamide in a PAGE gel was the major factor that influenced the relative position of the VP7 gene of G2 rotavirus strains (i.e., VP7 gene coding assignment by PAGE).
在各种轮状病毒重配体的开发和特性研究过程中,我们发现某些轮状病毒株编码中和及保护性抗原VP7的基因在聚丙烯酰胺凝胶电泳(PAGE)凝胶中的相对位置受丙烯酰胺浓度影响。
我们系统研究了影响轮状病毒VP7基因在PAGE凝胶中相对位置的各种因素。
我们通过在不同丙烯酰胺浓度下进行PAGE分析,对具有G1、G2、G3或G9特异性的选定轮状病毒株的双链RNA进行分析。
我们证明,三种G2株的VP7基因相对位置因PAGE凝胶中丙烯酰胺浓度而异,这种情况不仅发生在同源G2病毒基因背景中,也发生在异源G3病毒基因背景中;当改变PAGE电泳条件时,具有G1、G3、G4或G9特异性的VP7基因未表现出这种现象。
PAGE凝胶中丙烯酰胺的浓度是影响G2轮状病毒株VP7基因相对位置的主要因素(即通过PAGE进行VP7基因编码分配)。