HIGH-LEVEL EXPRESSION OF NONFUSED FOREIGN GENES WITH AUTOGRAPHA-CALIFORNICA NUCLEAR POLYHEDROSIS-VIRUS EXPRESSION VECTORS
HIGH-LEVEL EXPRESSION OF NONFUSED FOREIGN GENES WITH AUTOGRAPHA-CALIFORNICA NUCLEAR POLYHEDROSIS-VIRUS EXPRESSION VECTORS
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DOI：
10.1016/0042-6822(89)90348-6
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发表时间：
1989-05-01
期刊：
影响因子：
3.7
通讯作者：
SUMMERS, MD
中科院分区：
文献类型：
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作者：
LUCKOW, VA;SUMMERS, MD
高水平的非融合氯霉素乙酰转移酶，β-半乳糖苷酶和β-利用SDS-PAGE和RNA斑点杂交技术研究了多角体蛋白启动子控制下的葡萄糖醛酸酶在苜蓿银纹夜蛾核型多角体病毒感染的草地贪夜蛾（Spodoptera frugiperda）细胞中的表达。当多角体蛋白ATG起始密码子通过定点诱变转化为ATT时，在下游ATG密码子处开始翻译，导致高产量的非融合外源蛋白。当终止密码子插入下游和相位与多角体蛋白ATG密码子，但上游的ATG外源基因，非融合蛋白也产生，但在较低的水平。多角体蛋白基因启动的稳态mRNA水平不受ATG突变为ATT或在多角体蛋白ATG下游插入同相终止密码子的影响。
High levels of nonfused chloramphenicol acetyltransferase, .beta.-galactosidase, and .beta.-glucuronidase expressed under the control of new vector constructs of the polyhedrin promoter in Spodoptera frugiperda cells infected with Autographa californica nuclear polyhedrosis virus were investigated by SDS-PAGE and RNA dot blot analysis of total cytoplasmic RNA. When the polyhedrin ATG start codon was converted to ATT by site-directed mutagenesis, translation initiated at downstream ATG codons resulting in high yields of nonfused foreign proteins. When a stop codon was inserted downstream from and in phase with the polyhedrin ATG codon but upstream from the ATG of a foreign gene, nonfused proteins were also produced, but at lower levels. The levels of steady-state polyhedrin gene-promoted mRNA was not affected by the mutation from ATG to ATT or the insertion of in phase stop codons downstream from the polyhedrin ATG.