【摘要】 应用液氮冷冻香蕉束顶病组织,高速粉碎,氯仿—正丁醇澄清,差速离心,沉淀悬浮后经蔗糖垫部分纯化病毒,收集的蔗糖垫浓缩后经0—40％连续蔗糖梯度离心4小时,在离心管上半部形成一个明显的含有大量等径病毒的紫外吸收峰。再经不连续硫酸铯梯度离心6小时,形成一狭窄的乳白色带,含有高度纯化的直径约18nm的香蕉束顶病毒(BBTV)。用台湾提供的BBTV单克隆抗体,ELISA法可检测到提纯过程中病毒的存在,ISEM法可从粗提物中捕获大量等径病毒。ELISA法也用于检测田间及组织培养原种苗的带毒情况。更多还原

【Abstract】 Banana bunchy-top virus (BBTV) has been purified from diseased banana tissues in Fujian province,China. The banana tissues are pulverized with liquid nitrogen in a coffee grinder. The extracts are clarified with chloroform-butanol,and concentrated by one cycle of differential centrifugation. The suspension of the pellets is further centrifuged on the top of two layer sucrose cushions with 10% and 30% concentrations. The particles in the cushions are collected, concentrated, resuspended and centrifuged on the top of 0 - 40 % sucrose density gradient. The virus particles in a UV-absorbing band on the uppper tube of the sucrose density gradient are further purified in a discontinuous density gradient of cesium sulfate and form a narrow visible band containing a large number of highly purified isometric virus with the diameter about 18 nm. The virus is able to be detected in the process of purification by ELISA with the monoclonal antibody a-gainst BBTV which provided from Taiwan University, and be trapped by ISEM from the clarified extract. The monoclonal antibody has been used to diagnose the field-growing banana samples and tissue culture seedlings.更多还原