Ritter D G, Roy P
Environment Health Sciences Dept. University of Alabama, Birmingham.
Virus Res. 1988 Aug;11(1):33-47. doi: 10.1016/0168-1702(88)90065-2.
Full-length DNA clones representing the 10 double-stranded RNA segments of US bluetongue virus serotype 10 (BTV-10) have been used in a study to determine the genetic relationships among 20 different BTV serotypes. The study was undertaken using Northern blot hybridization techniques involving 32P-labelled DNA probes and total RNA species extracted from BHK-21 cells infected with 20 different BTV serotypes. The results obtained indicate that all the genes representing the nonstructural proteins of BTV (NS1, NS2 and NS3) as well as most of the inner capsid polypeptides are highly conserved (e.g., VP1, VP3, VP4), while VP6 and VP7, the remaining two inner capsid components, are less conserved. The genes representing the two outer capsid polypeptides, VP2 and VP5, vary significantly. When complete DNA clones of RNA segment 2 (representing the VP2 neutralization gene) of 4 other US serotypes (BTV-2, -11, -13 and -17) and one Australian serotype (BTV-1) were used in similar hybridization studies, the data obtained showed that despite geographical distances, a certain BTV serotype exhibits similarities. Some hybridization signals were detected with several of the inner capsid genes and the corresponding RNA segments of epizootic hemorrhagic disease virus (EHDV), a distantly related orbivirus, although none of the BTV outer capsid genes, nor any of the nonstructural genes hybridized with either EHDV-1 or EHDV-2 RNA species.
全长DNA克隆代表了美国蓝舌病病毒血清型10(BTV - 10)的10个双链RNA片段,已被用于一项研究中,以确定20种不同BTV血清型之间的遗传关系。该研究采用Northern印迹杂交技术,涉及用32P标记的DNA探针以及从感染20种不同BTV血清型的BHK - 21细胞中提取的总RNA。所得结果表明,代表BTV非结构蛋白(NS1、NS2和NS3)以及大多数内衣壳多肽的所有基因高度保守(例如VP1、VP3、VP4),而其余两个内衣壳成分VP6和VP7则保守性较低。代表两个外衣壳多肽VP2和VP5的基因差异显著。当使用其他4种美国血清型(BTV - 2、-11、-13和-17)和1种澳大利亚血清型(BTV - 1)的RNA片段2的完整DNA克隆(代表VP2中和基因)进行类似的杂交研究时,所得数据表明,尽管存在地理距离,某些BTV血清型仍表现出相似性。用几种内衣壳基因和流行性出血病病毒(EHDV,一种远缘相关的环状病毒)的相应RNA片段检测到了一些杂交信号,尽管BTV的任何外衣壳基因或任何非结构基因均未与EHDV - 1或EHDV - 2的RNA片段杂交。