| 对虾白斑病毒VP15基因的RNA干扰体外研究 |
| siRNA interference on VP15 gene of shrimp wssv |
| 投稿时间：2010-04-26 修订日期：2010-04-26 |
| DOI： |
| 中文关键词:WSSV VP15 RNA干扰 |
| 英文关键词:WSSV VP15 RNA interference |
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| 中文摘要: |
| ＶＰ１５是南美白对虾ＷＳＳＶ病毒的一个核衣壳蛋白基因。采用体外干扰实验筛选对ＶＰ１５具有针对性的ｓｉＲＮＡ。实验构建了真核表达载体ｐＥＧＦＰ－ＶＰ１５，并将设计的ｓｉＲＮＡ以及ｐＥＧＦＰ－ＶＰ１５共转染ＢＨＫ细胞。采用Ｗｅｓｔｅｒｎｂｌｏｔ方法检测ＧＦＰ－ＶＰ１５融合蛋白的表达，半定量ＲＴ－ＰＣＲ方法检验ｓｉＲＮＡ抑制ＶＰ１５基因转录的效果。实验结果表明，ｐＥＧＦＰ－ＶＰ１５在ＰＫ细胞正常表达，设计的３对ｓｉＲＮＡ对ＧＦＰ－ＶＰ１５的ｍＲＮＡ转录均有不同程度的干扰效果；其中，ｓｉＲＮＡ１的干扰效果最为显著。实验结果为下一步对虾体内干扰ＷＳＳＶ病毒复制研究以及筛选更多的特异ｓｉＲＮＡ建立基础。 |
| 英文摘要: |
| VP15，a structure gene for virion nucleocapsid. In order to screen out specific siRNA for VP15，an vitrovector named pEGFP-VP15 was constructed，and then siRNA targeting VP15 and pEGFP-VP15 were cotransfected into PK cell. The expression of pEGFP-VP15 was verified by Western blot，and the interference effect toVP15 in transcription was detected by semi-quantitative reverse transcription and polymerase Chain reaction. Our study here demonstrates that pEGFP-VP15 can be expressed in PK cell correctelly，the mRNA transcription for GFP-VP15 was suppressed by three pairs siRNA specifically，and the siRNAl made the most remarkable effect.Our study results would provide a system for choosing specifical siRNA for VP15 and also provide a foundation for further research such as suppressing the wssv reproduce in shrimp. |
| 黎铭,陈晓汉,马春霞,蒋伟明,马宁,熊建华,彭金霞,陈秀荔,彭敏,曾地刚,李咏梅.2010.对虾白斑病毒VP15基因的RNA干扰体外研究[J].水生态学杂志,31(5):56-61. |
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