论文部分内容阅读
葡萄A病毒(Grapevine virus A,GVA)是葡萄病毒属(Vitivirus)的典型种,在世界葡萄产区广泛分布。采集10株‘藤稔’葡萄成熟枝条,使用6种葡萄病毒ELISA试剂盒检测发现,10个样本中有6个感染4种不同的葡萄病毒。以GVA的ELISA阳性植株为材料进行RT-PCR扩增,首次获得了GVA四川分离物SL10的完整外壳蛋白基因(CP),该基因全长597bp。将其与GenBank收录的15个GVA分离物的CP序列进行比对和构建系统进化树,把不同地理起源的GVA分离物分成两个变异组,其中Ⅰ组包括3个分离物(与Ⅱ组的其他分离物只有75.9%~80.1%的序列同一性);其余的13个分离物组成Ⅱ组(组内分离物具有84.4%~99.5%的序列同一性)。构建了GVACP的原核表达质粒PET-30-GVAcp并转化BL21菌株,经IPTG诱导,目的基因得到了大量表达。
Grapevine virus A (GVA) is a typical genus of Vitivirus and is widely distributed in the world’s grapevines. Twenty mature grape shoots from ’Fujiminori’ were collected and tested using six grapevirus ELISA kits. Six out of 10 samples were found to infect 4 different grapevines. The complete coat protein gene (CP) of GVA Sichuan isolate SL10 was obtained for the first time by RT-PCR using GVA ELISA positive plants as the material, and the gene was 597 bp in length. The phylogenetic tree was constructed by comparing the CP sequences of the 15 GVA isolates collected from GenBank and the phylogenetic tree. The GVA isolates with different geographical origins were divided into two groups: Ⅰ group consisted of 3 isolates Other isolates only 75.9% to 80.1% sequence identity); the remaining 13 isolates form group II (isolates have 84.4% to 99.5% sequence identity). The prokaryotic expression plasmid PET-30-GVAcp of GVACP was constructed and transformed into BL21 strain. After induced by IPTG, the target gene was expressed in large quantities.