Use of a mammalian internal ribosomal entry site element for expression of a foreign protein by a transfectant influenza virus
Use of a mammalian internal ribosomal entry site element for expression of a foreign protein by a transfectant influenza virus
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使用哺乳动物内部核糖体进入位点元件通过转染流感病毒表达外源蛋白
作者：
A. García;T. Muster;W. Barclay;N. Percy;P. Palese
流感病毒核糖核蛋白转染系统使我们能够构建两种含有双顺反子神经氨酸酶（NA）基因的甲型流感病毒GP 2/BIP-NA和HGP 2/BIP-NA。来自双顺反子NA基因的mRNA具有两个不同的开放阅读框（ORF）。第一个ORF编码含有源自人免疫缺陷病毒1型gp 41蛋白的氨基酸序列的外源多肽（GP 2或HGP 2）。第二个ORF编码NA蛋白;它的翻译是通过一个内部核糖体进入位点实现的，该位点来源于人免疫球蛋白重链结合蛋白mRNA的5'非编码区。GP 2（79个氨基酸）和HGP 2（91个氨基酸）多肽在用相应的转染病毒感染的细胞中表达。HGP 2多肽包含与甲型流感/WSN/33病毒的血凝素（HA）蛋白相同的跨膜和胞质结构域，其被包装到病毒颗粒中。这种新的流感病毒系统涉及一个内部核糖体进入位点元件可能提供一种方法来表达各种外源基因在哺乳动物细胞。
The ribonucleoprotein transfection system for influenza virus allowed us to construct two influenza A viruses, GP2/BIP-NA and HGP2/BIP-NA, which contained bicistronic neuraminidase (NA) genes. The mRNAs derived from the bicistronic NA genes have two different open reading frames (ORFs). The first ORF encodes a foreign polypeptide (GP2 or HGP2) containing amino acid sequences derived from the gp41 protein of human immunodeficiency virus type 1. The second ORF encodes the NA protein; its translation is achieved via an internal ribosomal entry site which is derived from the 5' noncoding region of the human immunoglobulin heavy-chain-binding protein mRNA. The GP2 (79 amino acids) and HGP2 (91 amino acids) polypeptides are expressed in cells infected with the corresponding transfectant virus. The HGP2 polypeptide, which contains transmembrane and cytoplasmic domains identical to those of the hemagglutinin (HA) protein of influenza A/WSN/33 virus, is packaged into virus particles. This novel influenza virus system involving an internal ribosomal entry site element may afford a way to express a variety of foreign genes in mammalian cells.