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首页> 外文期刊>Journal of Fish Diseases >Viral susceptibility, transfection and growth of SPB - a fish neural progenitor cell line from the brain of snubnose Domoano. Trachinotus blochii ILaceoede)
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Viral susceptibility, transfection and growth of SPB - a fish neural progenitor cell line from the brain of snubnose Domoano. Trachinotus blochii ILaceoede)

机译:病毒易感性,转染和SPB的生长-SPB是一种来自蛇形Domoano脑的鱼类神经祖细胞系。 Trachinotus blochii ILaceoede)

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This study investigates the susceptibilities of the SPB cell line to fish viruses including giant sea-perch iridovirus (GSIV-K1), red sea bream iridovirus (RSIV-Ku), grouper nervous necrosis virus (GNNV-K1), chum salmon reovirus (CSV) and eel herpesvirus (HVA). GSIV-K1, RSIV-Ku and CSV replicated well in SPB cells, with a significant cytopathic effect and virus production. However, the cells were HVA and GNNV refractory. To examine the ability of SPB cells to stably express foreign protein, expression vectors encoding GNNV Bl and B2 fused to enhanced green fluorescent protein (EGFP) and GSIV ORF35L fused to DsRed were constructed and introduced by transfection into SPB cells. Stable transfectants displayed different morphologies compared with SPB and with each other. EGFP-B1 was predominantly localized in the nuclei, EFPF-B2 was distributed throughout the cytoplasm and nucleus, and granular 35L-DsRed was localized with secreted vesicles. The expression of EFPF-B2 in SPB cells produced blebs on thesurface, but the cells showing stable expression of EGFP, EGFP-B1 or 35L-DsRed showed normal morphologies. Results show the SPB cells and the transfected cells grow well at temperatures between 20 and 35 °C and with serum-dependent growth. SPB cells aresuitable for studies on foreign protein expression and virology.
机译:这项研究调查了SPB细胞系对鱼类病毒的敏感性,其中包括巨型海鲈虹膜病毒(GSIV-K1),红鲷虹膜病毒(RSIV-Ku),石斑鱼神经坏死病毒(GNNV-K1),鲑鲑呼肠孤病毒(CSV) )和鳗鱼疱疹病毒(HVA)。 GSIV-K1,RSIV-Ku和CSV在SPB细胞中复制良好,具有明显的细胞病变作用和病毒产生。但是,这些细胞是HVA和GNNV难治性的。为了检查SPB细胞稳定表达外源蛋白的能力,构建了编码与增强的绿色荧光蛋白(EGFP)融合的GNNV B1和B2以及与DsRed融合的GSIV ORF35L的表达载体,并通过转染导入SPB细胞中。与SPB和彼此相比,稳定的转染子显示出不同的形态。 EGFP-B1主要位于细胞核中,EFPF-B2分布在整个细胞质和细胞核中,而颗粒状的35L-DsRed则位于分泌的囊泡中。 EFPF-B2在SPB细胞中的表达在表面产生了气泡,但显示EGFP,EGFP-B1或35L-DsRed稳定表达的细胞表现出正常的形态。结果显示,SPB细胞和转染的细胞在20至35°C的温度下生长良好,并具有血清依赖性生长。 SPB细胞适合用于外源蛋白表达和病毒学研究。

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