对虾白斑综合征病毒ORF220基因在昆虫细胞内的表达及其分布

对虾白斑综合征病毒厦门分离株ORF220编码真核生物GP130受体同源蛋白。将ORF220和绿色荧光蛋白编码基因融合在一起克隆到昆虫杆状病毒表达载体pFastBacI，然后与AcBacmid共同转染DH10B细胞。用PCR鉴定含有ORF220和EGFP基因的重组质粒，提取纯化重组质粒并转染昆虫细胞进行表达。结果发现，DNA转染后3-5d可以在荧光显微镜下观察到绿色荧光，表明融合蛋白在昆虫系统内成功表达。用病毒上清液感染昆虫细胞进行时相观察，结果表明，ORF220蛋白在昆虫细胞的细胞质和细胞核内呈随机分布，没有特异的细胞定位。

Expression and Distribution of White Spot Syndrome Virus ORF220 Gene in Insect Cells

White spot syndrome virus ORF220 encodes a protein homologous to the gp130 receptors of eukaryote. prcvious studies showed that this protein was a minor structural protein. In this paper, the ORF220 was cloned into the pFastBacI fused with EGFP gene and co-transfected with AcBacmid into DH10B cells. The recombinant Bacmid containing fused ORF220 and EGFP genes was confirmed by PCR and then transfected into Sf21 cells. The green fluorencence was observed 3-5 d post-transfection under the fluorescent microscopy and this confirmed that the fusion protein was expressed in insect cells. The supernatant of the transfected cells was used to infect the Sf21 cells and the time course observation showed that the ORF220 product localized in both the cytoplasm and nucleus with no predilection.