对虾一种杆状病毒多角体蛋白基因的PCR扩增

根据已发表的几种杆状病毒的多角体基因的序列比较与分析，设计并合成两对ＰＣＲ简并引物Ｐ—６０／Ｐ７４０和Ｐ１６４／Ｐ６４０，从纯化的对虾皮下及造血组织坏死杆状病毒提取ＤＮＡ并进行ＰＣＲ扩增，Ｐ—６０／Ｐ７４０的扩增片段较为复杂，基中１条的长度为８００ｂｐ的主带与设计的ＤＮＡ片段长度相近。Ｐ１６４／Ｐ６４０经优化ＰＣＲ的反应条件后，成功地扩增出与设计相同的约４８０ｂｐＤＮＡ片段。进一步的实验表明，Ｐ—６０／Ｐ７４０扩增的分子量为８００ｂｐ的片段可被Ｐ１６４／Ｐ６４０引物对扩增，分子量与预期的相同，初步研究结果显示，Ｐ—６０／Ｐ７４０可用于杆状病毒多角体蛋白全基因的克隆，Ｐ１６４／Ｐ６４０对于对虾杆状病毒的调查和研究有实际应用价值。

PRIMARY STUDIES ON THE POLYHEDRIN GENE OF AN BACULOVIRUS ISOLATED FROM PENAEID SHRIMP

Based on publicated sequences,two pairs of PCR primers, P-60/P740 and P164/P640,were designed and synthesized for the polyhedrin gene of baculovirus.The DNA extracted from purified hypodermal and hematopoietic necrosis baculovirus(HHNBV)was used as template and the expected PCR products were amplified. Though the pattern of P-164/P740 products were not singular, one of the main bands, approximately 800bp in length, tallied with designation. A specific DNA band,480bp in length was produced by P164/P640 and the size of this fagment conformed to expectation. Further investigation proved that the purified 800bp PCR product of P-60/P740 could be amplified by P164/P640 and the product was also 480bp in length. p-60/p740 were proposed to be used to clone the complete polyhedrin gene and P164/P640 to detect the baculovirus in shrimp culture.