Cloning, characterization, and phylogenetic analysis of a shrimp white spot syndrome virus gene that encodes a protein kinase
Cloning, characterization, and phylogenetic analysis of a shrimp white spot syndrome virus gene that encodes a protein kinase
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DOI：
10.1006/viro.2001.1091
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发表时间：
2001-10-25
期刊：
影响因子：
3.7
通讯作者：
Kou, GH
中科院分区：
文献类型：
--
作者：
Liu, WJ;Yu, HT;Kou, GH
在对虾白色斑点综合征病毒（WSSV）基因组的8421 bp EcoR 1片段中发现一个开放阅读框（ORF），编码715个氨基酸的多肽。该多肽与真核生物丝氨酸/苏氨酸蛋白激酶（PK）具有显著的同源性，并且包含真核生物蛋白激酶的主要保守亚结构域。偶联在体外转录和翻译产生的蛋白质具有约87 kDa的表观分子量，根据十二烷基硫酸钠-聚丙烯酰胺凝胶电泳。为进行pk基因的转录分析，在感染后不同时间从WSSV感染的虾中分离总RNA。北方印迹分析与pk特异性核糖核酸探针发现一个主要和次要的转录本的2.7和5.7 kb，分别。快速扩增2.7kb pk主要转录本的5 ′端cDNA末端，发现在ATG翻译起始密码子的-38（G）和-39（G）核苷酸残基处有两个转录起始位点。RT-PCR分析表明，pk基因在感染后2 h开始转录，并持续至少60 h。系统发育分析表明，WSSV蛋白激酶没有任何近亲，不属于任何主要的蛋白激酶组。(C)北京：科学出版社.
An open reading frame (ORF) that encodes a 715-amino-acid polypeptide was found in an 8421-bp EcoR1 fragment of the shrimp white spot syndrome virus (WSSV) genome. The polypeptide shows significant homology to eukaryotic serine/threonine protein kinase (PK) and contains the major conserved subdomains for eukaryotic protein kinases. Coupled in vitro transcription and translation generated a protein having an apparent molecular mass of about 87 kDa according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. For transcriptional analysis of the pk gene, total RNA was isolated from WSSV-infected shrimp at different times after infection. Northern blot analysis with pk-specific riboprobe found a major and a minor transcript of 2.7 and 5.7 kb, respectively. Rapid amplification of the 5' cDNA ends of the major 2.7-kb pk transcript showed that there were two transcriptional initiation sites located at nucleotide residues -38(G) and -39(G) relative to the ATG translational start codon. Temporal expression analysis by RT-PCR indicated that the transcription of the pk gene started 2 h after infection and continued for at least 60 h. Phylogenetic analysis showed that WSSV protein kinase does not have any close relatives and does not fall into any of the major protein kinase groups. (C) 2001 Academic Press.