BACULOVIRUS EXPRESSION VECTORS - THE REQUIREMENTS FOR HIGH-LEVEL EXPRESSION OF PROTEINS, INCLUDING GLYCOPROTEINS
BACULOVIRUS EXPRESSION VECTORS - THE REQUIREMENTS FOR HIGH-LEVEL EXPRESSION OF PROTEINS, INCLUDING GLYCOPROTEINS
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DOI：
10.1099/0022-1317-68-5-1233
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发表时间：
1987-05-01
影响因子：
3.8
通讯作者：
BISHOP, DHL
中科院分区：
文献类型：
--
作者：
MATSUURA, Y;POSSEE, RD;BISHOP, DHL
研究了利用银纹夜蛾核型多角体病毒多角体基因启动子高效表达3种外源蛋白的条件。在感染适当重组杆状病毒的果蝇细胞中，淋巴细胞性脉络膜脑膜炎病毒(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。当所有多角体基因编码序列或部分紧邻的3‘’下游序列被删除时，对N蛋白的表达水平没有影响。利用从新的转移载体pacYMI获得的最有效的表达病毒，估计LCMV N蛋白约占细胞总蛋白的50%，这与感染细胞的细胞质中存在大量包涵体是一致的。从含有LCMV GPC基因的pAcYM1转移载体获得的重组病毒，其糖蛋白的合成水平约相当于细胞蛋白的20%。感染GPC重组体的细胞薄片显示高度空泡化的细胞质。
The requirements for high level expression of three foreign proteins using the polyhedrin gene promoter of Autographa californica nuclear polyhedrosis virus (AcNPV, Baculoviride) 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 efficent expression viruses derived from a new transfer vector, pacYMI, 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.