CAJ | 학술논문

以感染锦鲤疱疹病毒的患病鱼为材料,通过RT-PCR克隆ORF134的编码序列,在细胞水平分析其表达模式,并在原核表达系统中表达,获得可溶性的ORF134重组蛋白。结果表明ORF134在锦鲤疱疹病毒裂解感染中转录,证实ORF134为非结构功能基因。 ORF134全长为624 bp,含有1个内含子,编码179个氨基酸,序列分析显示ORF134同已报道的高等脊椎动物病毒编码的vIL-10具有较低的序列同源性,但具有较高的结构相似性。体外表达分析显示ORF134呈分泌性表达,但不能形成寡聚体,与其他病毒编码的vIL-10具有差异。进一步通过原核表达获得可溶性的ORF134重组蛋白。
이감염금리포진병독적환병어위재료,통과RT-PCR극륭ORF134적편마서렬,재세포수평분석기표체모식,병재원핵표체계통중표체,획득가용성적ORF134중조단백。결과표명ORF134재금리포진병독렬해감염중전록,증실ORF134위비결구공능기인。 ORF134전장위624 bp,함유1개내함자,편마179개안기산,서렬분석현시ORF134동이보도적고등척추동물병독편마적vIL-10구유교저적서렬동원성,단구유교고적결구상사성。체외표체분석현시ORF134정분비성표체,단불능형성과취체,여기타병독편마적vIL-10구유차이。진일보통과원핵표체획득가용성적ORF134중조단백。
Cyprinid herpesvirus 3 ( CyHV-3) , also known as koi herpesvirus ( KHV) , is the causative a-gent of an emerging and mortal disease in common ( Cyprinus carpio carpio) and koi ( Cyprinus carpio koi) carp. Since its emergence in the late 1990s, KHV has caused severe financial and economic losses in both koi and common carp culture industries worldwide. CyHV-3 contains an interleukin-10 like open reading frame (ORF134), which has been identified from multiple members of Herpesvirales and Poxviridae. Here in this study, ORF134 of CyHV-3 was amplified by RT-PCR. It is 624 bp in full-length with an intron and encodes a peptide of 179 amino acids. Multiple sequence alignment indicated that ORF134 shared 23. 5% amino acid se-quence similarity with carp IL-10 . Homology modeling suggested that ORF134 had a similar three-dimensional structure with vIL-10 encoded by EBV, but shared low sequence similarity with other vIL-10. To analysis the ex-pression profile of ORF134 , the coding region of ORF134 was cloned into pcDNA3 . 1 vector and expressed in 293T cells. The results suggested that ORF134 could secrete from expressed cells, but different from other iden-tified viral IL-10 homologs, ORF134 could not form homodimer under non-reduced condition. Further study showed that soluble fusing protein of ORF134 was also expressed in E. coli.