【作者】 杨国慧； 崔崇士； 张仲凯；

【Author】 YANG Guo-hui~1,CUI Chong-shi~1,ZHANG Zhong-kai~2 (1 Horticultural College of Northeast Agricultural University,Harbin 150030,China;2 Yunnan Key Lab of Agro-Biotechnology;Kunming 650223,China)

【机构】 东北农业大学园艺学院； 云南省农业生物技术重点实验室；

【摘要】 利用电镜和酶联免疫吸附测定法(ELISA)在黑龙江省采集的南瓜病样中检测到西瓜花叶病毒2号(WMV-2)。再利用免疫PCR(IC-PCR)和反转录PCR(RT-PCR)方法,扩增获得其外壳蛋白(CP)基因片段,并克隆到pGEM-T载体中。核苷酸序列测定表明,该分离物CP基因全长为852个核苷酸,编码由284个氨基酸组成的31.8 kDa蛋白。与国外已报道的WMV-2 CP基因相比,其核苷酸序列同源性为92.2%-94.0%,由此推导的氨基酸序列同源性为94.5%-98.1%。与国内2个分离物相比,和山西分离物核苷酸和氨基酸的同源性都达到98.5%,和郑州分离物核苷酸和氨基酸的同源性分别为91.5%和95.0%。更多还原

【Abstract】 Watermelon mosaic virus 2(WMV-2) was detected in infected pumpkin plant collected from Heilongjiang(HLJ) province by electron microscopy and enzyme-linked immunosorbent assay(ELISA).A specific fragment about 850 bp in length covering coat protein (CP) gene region was amplified by reverse transcription polymerase chain reaction(RT-PCR) and immuno capture PCR(IC-PCR).The PCR product was cloned into pGEM-T vector. Sequences analysis showed that CP gene of WMV-2 isolate HLJ had 852 nucleotides in length, encoding for the coat protein with molecular weights of 31.8 Kda.Compared with the published sequences abroad,it shared identities of 92.2-94.0%in nucleotide acids and 94.5-98.1%in amino acid respectively.Compared with WMV-2-Shanxi and WMV-2-Zhengzhou CP gene sequences of China,the homology was 98.5%,91.5%in nucleotide acids and 98.5%,95.0%in amino acids respectively.更多还原