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小鹅瘟病毒结构蛋白的分析与 PCR 鉴定
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李永明，郑大恒，温贵兰，文 明
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（贵州大学 动物科学学院，贵州 贵阳 550025）
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摘要：
采用聚乙二醇（PEG 6000）沉淀和Sepharose 4B层析法，从感染小鹅瘟病毒的鹅胚尿囊液浓缩和纯化了小鹅瘟病毒，经测定其核酸长度为5 000 bp，含3条结构蛋白带，VP1、VP2和VP3的分子质量依次为88、68和57 ku，其中VP3是病毒的主要结构蛋白。针对编码VP3的基因设计合成1对引物，以小鹅瘟病毒感染鹅胚尿囊液为反应模板，PCR扩增出了1条530 bp的片段，与预 期设计的长度基本相符；Ssp Ⅰ酶切分析表明，扩增产物是目的基因片段;这对引物能够特异性地扩增小鹅瘟病毒的核酸，与其他禽类病毒核酸不发生交叉反应。 |
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关键词：
小鹅瘟病毒；核酸；结构蛋白；多聚酶链反应；酶切分析 |
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中图分类号：S 852.659.2 文献标识码：
A 文章编号：1000-6419(2005)S1-0013-04 |
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Structural protein analysis and PCR identification of gosling plague virus |
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LI Yong-ming，ZHENG Da-heng，WEN Gui-lan，WEN Ming |
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College of Animal Science，Guizhou University，Guiyang 550025，China）
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Abstract: The gosling plague virus had been concentrated and purified from allantoic fluid of affected embryos by PEG 6000 precipitation and Sepharose 4B chromatography. The length of the virus nucleic acid was determined as 5 000 bp. Three bands of structural protein were demonstrated with the molecular weight of 88 ku（VP1），68 ku（VP2）and 57 ku（VP3）. Among them VP3 was the main structural protein of the virus. A pair of primers has been designed and synthesized，which was directed to the gene coded for VP3. When the allantoic fluid of affected embryos was used as template，a fragment of 530 bp had been amplified by PCR，Which was in accordance with the expected length. The restriction endonuclease analy-sis of Ssp Ⅰ indicated that the PCR product was the target gene fragment. The primers could specifically amplified the nucleic acid of gosling plague virus and no cross reaction was observed with other avian viru-ses. |
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Key words: gosling plague virus；nucleic acid；structural protein；polymerase chain reaction；restric-tion endonuclease analysis |
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