Harper G, Dahal G, Thottappilly G, Hull R
Department of Virus Research, John Innes Centre, Norwich, Norfolk, UK.
J Virol Methods. 1999 Apr;79(1):1-8. doi: 10.1016/s0166-0934(98)00158-x.
A polymerase chain reaction (PCR) based strategy to detect episomal banana streak badnavirus (BSV) in banana and plantain plants that carry integrated BSV sequences was developed. Antisera used in immuno-capture polymerase chain reaction (IC-PCR) are capable of binding a large number of BSV serotypes. The primers used for PCR are capable of annealing to and amplifying across the aspartic protease-reverse transcriptase domain boundaries of both episomal and integrated BSV sequences and result in similar or identical sequence size fragments from either template. However, we show that under the conditions selected for IC-PCR, nuclear, mitochondrial or chloroplast genomic sequences are not amplified and thus only captured episomal BSV is amplified. IC-PCR is suitable for the large-scale screening of Musa for episomal BSV which is necessary for germplasm movement.
开发了一种基于聚合酶链反应(PCR)的策略,用于检测携带整合型香蕉条纹杆状病毒(BSV)序列的香蕉和大蕉植株中的游离型香蕉条纹杆状病毒(BSV)。免疫捕获聚合酶链反应(IC-PCR)中使用的抗血清能够结合大量的BSV血清型。用于PCR的引物能够与游离型和整合型BSV序列的天冬氨酸蛋白酶-逆转录酶结构域边界退火并进行扩增,并且从任一模板产生大小相似或相同的序列片段。然而,我们表明,在为IC-PCR选择的条件下,核、线粒体或叶绿体基因组序列不会被扩增,因此只有捕获的游离型BSV会被扩增。IC-PCR适用于大规模筛选香蕉中的游离型BSV,这对于种质转移是必要的。