Wang Yong-Jie, Chen Ke-Ping, Yao Qin, Han Xu
Institute of Fisheries, Anhui Academy of Agricultural Sciences, Hefei 230031, P. R. China.
J Microbiol. 2007 Apr;45(2):139-45.
The Bombyx mori parvo-like virus (China isolate) DNA polymerase (BmDNV-3 dnapol) gene has been tentatively identified based on the presence of conserved motifs. In the present study, we perform a transcriptional analysis of the BmDNV-3 dnapol gene using the total RNA isolated from BmDNV-3 infected silkworm at different times. Northern blot analysis with a BmDNV-3 dnapol-specific riboprobe showed a major transcript of 3.3 kb. 5'-RACE revealed that the major transcription start point was located 20 nucleotides downstream of the TATA box. In a temporal expression analysis using differential RT-PCR, BmDNV-3 dnapol transcript was detected at low levels at 6 h.p.i., increased from 6 to 36 h.p.i., and remained fairly constant thereafter. Analysis of the predicted DNA polymerase sequence using neighborjoining and protein parsimony algorithms indicated that the predicted 1115-residue polypeptide contained five motifs associated with DNA polymerases synthetic activities and three additional motifs associated with polymerases possessing 3' to 5' exonuclease activity. The molecular phylogenetic analysis of this gene supported the placement of Bombyx mori parvo-like virus in a separate virus family.
基于保守基序的存在,家蚕细小病毒样病毒(中国分离株)DNA聚合酶(BmDNV-3 dnapol)基因已被初步鉴定。在本研究中,我们使用从不同时间感染BmDNV-3的家蚕中分离的总RNA对BmDNV-3 dnapol基因进行转录分析。用BmDNV-3 dnapol特异性核糖探针进行的Northern印迹分析显示有一个3.3 kb的主要转录本。5'-RACE显示主要转录起始点位于TATA框下游20个核苷酸处。在使用差异RT-PCR的时间表达分析中,BmDNV-3 dnapol转录本在感染后6小时检测到低水平,从感染后6小时到36小时增加,此后保持相当恒定。使用邻接法和蛋白质简约算法对预测的DNA聚合酶序列进行分析表明,预测的1115个残基的多肽包含与DNA聚合酶合成活性相关的五个基序以及与具有3'至5'核酸外切酶活性的聚合酶相关的另外三个基序。该基因的分子系统发育分析支持将家蚕细小病毒样病毒置于一个单独的病毒科中。