Jose Jency, Jalali S K, Shivalingaswamy T M, Kumar N K Krishna, Bhatnagar R, Bandyopadhyay A
Molecular Entomology Laboratory, National Bureau of Agriculturally Important Insects, Post Bag No. 2491, H. A. Farm Post, Bellary Road, Hebbal, Bangalore, 560024 Karnataka India.
International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi, 110067 India.
Indian J Virol. 2013 Jun;24(1):59-65. doi: 10.1007/s13337-013-0126-3. Epub 2013 Jan 29.
A PCR based method for detection of viral DNA in nucleopolyhedrovirus of three lepidopterans, Spodoptera litura, Amsacta albistriga and Helicoverpa armigera, was developed by employing the late expression factor-8 (lef-8) gene of three NPV using specific primers. The amplicons of 689, 699 and 665 bp were amplified, respectively, and the nucleotide sequences were submitted to GenBank and the accession numbers were obtained. The sequences of lef-8 gene of S. litura NPV and H. armigera NPV matched with those of their respective references in the GenBank database, thereby confirming their identity, however, the sequence of A. albistriga NPV was the first sequence submitted to the GenBank database. The sequence similarity analysis between the three lef-8 gene of NPV sequenced in the present study revealed that there was no significant similarity between them, however A. albistriga NPV and S. litura NPV were found to be closely related. CLUSTAL alignment of the sequences generated revealed general relatedness among NPVs lef-8 gene. The study confirmed that lef-8 gene can be used for quick and correct discriminatory identification of insect viruses.
通过使用特异性引物,利用三种核型多角体病毒(NPV)的晚期表达因子8(lef-8)基因,开发了一种基于PCR的方法,用于检测三种鳞翅目昆虫斜纹夜蛾、白纹污灯蛾和棉铃虫的核型多角体病毒中的病毒DNA。分别扩增出689、699和665 bp的扩增子,并将核苷酸序列提交至GenBank并获得登录号。斜纹夜蛾NPV和棉铃虫NPV的lef-8基因序列与其在GenBank数据库中的各自参考序列匹配,从而确认了它们的身份,然而,白纹污灯蛾NPV的序列是首次提交至GenBank数据库的序列。本研究中测序的三种NPV的lef-8基因之间的序列相似性分析表明,它们之间没有显著的相似性,然而发现白纹污灯蛾NPV和斜纹夜蛾NPV密切相关。对所产生序列的CLUSTAL比对揭示了NPV的lef-8基因之间的一般相关性。该研究证实lef-8基因可用于快速、准确地鉴别昆虫病毒。