Matsuura Y, Possee R D, Overton H A, Bishop D H
J Gen Virol. 1987 May;68 ( Pt 5):1233-50. doi: 10.1099/0022-1317-68-5-1233.
The requirements for high level expression of three foreign proteins using the polyhedrin gene promoter of Autographa californica nuclear polyhedrosis virus (AcNPV, Baculoviridae) have been investigated. In Spodoptera frugiperda cells infected with the appropriate recombinant baculoviruses, the synthesis of the two S RNA coded genes of lymphocytic choriomeningitis virus (LCMV; i.e. the nucleoprotein, N, and glycoprotein precursor, GPC), or the haemagglutinin gene of influenza A virus, appears to be related to the degree of integrity of the 5' upstream sequence of the polyhedrin gene. No effect on the level of N protein expression was detected when all the polyhedrin gene coding sequences or some of the immediate 3' downstream sequences were deleted. Using the most efficient expression viruses derived from a new transfer vector, pAcYM1, it has been estimated that LCMV N protein represented approximately 50% of the total cellular protein, an observation consistent with the presence of numerous inclusion bodies in the cytoplasm of infected cells. For recombinant viruses derived from the pAcYM1 transfer vector containing the LCMV GPC gene, the level of synthesis of the arenavirus glycoprotein was equivalent to approximately 20% of the cellular protein. Thin sections of cells infected with the GPC recombinant revealed a highly vacuolated cytoplasm.
利用苜蓿银纹夜蛾核型多角体病毒（AcNPV，杆状病毒科）的多角体蛋白基因启动子高水平表达三种外源蛋白的条件已得到研究。在被合适的重组杆状病毒感染的草地贪夜蛾细胞中，淋巴细胞性脉络丛脑膜炎病毒（LCMV）的两个S RNA编码基因（即核蛋白N和糖蛋白前体GPC）或甲型流感病毒的血凝素基因的合成，似乎与多角体蛋白基因5'上游序列的完整性程度有关。当删除所有多角体蛋白基因编码序列或一些紧邻的3'下游序列时，未检测到对N蛋白表达水平的影响。使用源自新转移载体pAcYM1的最有效的表达病毒，据估计LCMV N蛋白约占细胞总蛋白的50%，这一观察结果与感染细胞胞质中存在大量包涵体一致。对于源自含有LCMV GPC基因的pAcYM1转移载体的重组病毒，沙粒病毒糖蛋白的合成水平约相当于细胞蛋白的20%。用GPC重组体感染的细胞的超薄切片显示出高度空泡化的细胞质。
