湖北省柑桔裂皮类病毒的检测及序列分析

以表现柑桔裂皮病典型症状的罗伯逊脐橙为材料,采用双向电泳、RT-PCR和RNA斑点杂交及组织印迹杂交的方法,对柑桔裂皮类病毒进行了检测,结果表明RT-PCR和2种核酸杂交技术均可用于该类病毒的有效检测。对该类病毒全长RNA的RT-PCR产物进行了克隆和序列分析,该分离株(命名为CEVd-HB分离株)的全长RNA由371个核苷酸组成,与已报道的其它不同来源分离株相似性为90%~99%。     

香蕉线条病毒ORFⅡ基因的原核表达及抗体制备

 ORFⅡ gene of Banana streak virus GuangDong isolate (BSV-GD) was amplified from a BSV-GD recombinant plasmid by PCR, and the gene was expressed by being cloned into prokaryote expression vector pET-28b (+). The fusion protein was about 16.5 kDa in size and was soluble with SDS-PAGE analysis. The purified protein was obtained by using the histidine labeling kit of N-terminus of protein. The antiserum was obtained by immunizing healthy rabbits with the purified protein. Western blot and ELISA analysis showed that the special antiserum of BSV possessed high titer, which was tested as 1:51 200. The study was a base for further research on BSV including ORFⅡ gene function and virus detection.     

柑桔裂皮类病毒研究进展

<正>柑桔裂皮病是柑桔的主要病害之一,严重影响世界范围内柑桔的生产。起初认为柑桔裂皮病是由病毒引起的,但是进一步的研究表明该病的病原是类病毒。柑桔裂皮类病毒(citrusexocortis viroid,CEVd)属于马铃薯纺锤块茎类病毒属(Pospiviroid),大小一般为370-375nt(核苷酸),1994年Semancik等报道了一个大小为463 nt的该类病毒突变株,具有棍棒状二级结构。它的热钝化温度高,110℃下保持10-15分钟,仍不丧失其侵染力,到140℃热致死点时,感染性才呈线性下降。     

香蕉线条病毒ORF Ⅰ基因的原核表达及抗体制备

利用PCR方法从含香蕉线条病毒广东分离物(BSV-GD)部分基因组的质粒中扩增该病毒的ORF Ⅰ基因,将其克隆到pET-28b(+)原核表达载体中进行了融合表达.SDS-PAGE电泳发现,目的融合蛋白与预期大小一致,相对分子质量约为23 000,表达产物主要以不可溶的包涵体形式存在,将其变性溶解后利用N端的组氨酸标签进行纯化.以纯化产物为抗原免疫兔子制备了香蕉线条病毒的阳性兔抗血清,Western blot和ELISA分析表明,制备的兔抗血清为香蕉线条病毒的高效特异性抗血清,效价高达1:51 200.