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当前位置: 首页 > 文章 > 阴离子交换层析分离鲍肌肉萎缩症病毒 南方农业学报 2015,46 (12) 2223-2228
Position: Home > Articles > Isolation of abalone shriveling syndrome-associated virus by anion exchanger chromatography Journal of Southern Agriculture 2015,46 (12) 2223-2228

阴离子交换层析分离鲍肌肉萎缩症病毒

作  者：
谷露;姜敬哲;王江勇
单  位：
中国水产科学研究院南海水产研究所/农业部南海渔业资源开发利用重点实验室
关键词：
鲍;肌肉萎缩症病毒;分离;纯化;阴离子交换层析;电导率
摘  要：
【目的】建立与优化阴离子交换层析分离纯化鲍肌肉萎缩症病毒(Abalone shriveling syndrome-associated virus,Ab SV)的方法,为今后深入研究其形态结构及致病性打下基础。【方法】利用病毒表面带负电的特性,建立Q Sepharose Fast Flow阴离子交换层析分离鲍组织中Ab SV的方法,同时结合荧光定量PCR检测Ab SV含量以确定其流出峰和电导率。【结果】Ab SV峰集中在电导率为32~40 mg/cm时所收集的组分中。将收集液用30 k D滤膜浓缩后进行SDS-PAGE凝胶电泳检测,发现Ab SV阳性样品呈现出特有的蛋白条带。对特异蛋白条带切胶回收后进行质谱分析,证实5条蛋白条带中有2条鉴定为Ab SV重组酶。【结论】以阴离子交换层析分离鲍肌肉萎缩症病毒具有可行性,其电导率为32~40 mg/cm时的流出峰为Ab SV组分。
译  名：
Isolation of abalone shriveling syndrome-associated virus by anion exchanger chromatography
作  者：
GU Lu;JIANG Jing-zhe;WANG Jiang-yong;South China Sea Fisheries Research Institute,China Academy of Fishery Sciences,Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization,Ministry of Agriculture;College of Fisheries and Life,Shanghai Ocean University;
关键词：
abalone;;shriveling syndrome-associated virus;;isolate;;purify;;anion exchanger chromatography;;conductivity
摘  要：
【Objective】The anion exchanger chromatography technology was established and optimized to isolate and purify abalone shriveling syndrome-associated virus(Ab SV), in order to lay the foundation for further study of its morphological structure and pathogenicity. 【Method】Taking advantage of electronegativity on virus surface, Q Sepharose Fast Flow anion exchanger chromatography technology was utilized to isolate Ab SV from abalone. Then the Ab SV content of isolated sample was detected by real-time fluorescent quantitative PCR to confirm eluting peak and conductivity of Ab SV. 【Result】The results showed that the Ab SV eluent was isolated and collected when the conductivity was 32-40 mg/cm. And the collected Ab SV eluent was concentrated with 30 k D filter membrane, then detected by SDS-PAGE. The SDS-PAGE result showed that some specific protein bands were found to be in the Ab SV-positive sample. Furthermore, the specific bands were extracted from excised gels, and identified by mass spectrometry analysis, The identification result showed that two of five protein bands were identified as recombinase of Ab SV. 【Conclusion】It is feasible to use anion exchanger chromatography technology to isolate Ab SV, and the component of outflow peak is Ab SV at the conductance of 32-40 mg/cm.

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