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首页> 外文期刊>Protein Expression and Purification >IMMEDIATE-EARLY BACULOVIRUS VECTORS FOR FOREIGN GENE EXPRESSION IN TRANSFORMED OR INFECTED INSECT CELLS
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IMMEDIATE-EARLY BACULOVIRUS VECTORS FOR FOREIGN GENE EXPRESSION IN TRANSFORMED OR INFECTED INSECT CELLS

机译:用于转化或感染的昆虫细胞中外源基因表达的早期杆状病毒载体

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Baculovirus expression vectors are used routinely for foreign gene expression and are under intense development as improved biological pesticides. Conventional baculovirus expression vectors are recombinant viruses that can express a foreign gene in insect cells under the control of the polyhedrin promoter, which provides high-level transcription during the very late phase of infection. For some applications, including foreign glycoprotein production and insect pest control, it might be advantageous to have baculovirus vectors that could express foreign gene products in uninfected cells or earlier after infection. To fulfill this need, we have constructed a new set of plasmids that can be used to clone and express foreign genes under the control of a baculovirus iel promoter, which is active in uninfected insect cells and throughout infection. We used a subset of these new plasmids to isolate recombinant baculoviruses containing various foreign genes and compared expression of these genes by the resulting immediate-early baculovirus vectors and by conventional baculovirus vectors. As expected, the immediate-early vectors began to express each foreign gene earlier in infection but, by 36-48 h postinfection, the conventional vectors had produced more of each foreign protein. Conventional baculovirus vectors also produced more enzymatic activity from two different procaryotic genes than the immediate-early baculovirus vectors. However, immediate-early vectors produced as much or more enzymatic activity from two different eucaryotic genes encoding secretory pathway proteins than the conventional vectors, even at 48 h postinfection. Hence, this report describes a new set of plasmids that can be used to clone and express foreign genes under the control of the baculovirus iel promoter and suggests that immediate-early baculovirus vectors might be as useful as conventional baculovirus expression vectors for producing biologically active eucaryotic secretory pathway proteins. (C) 1996 Academic Press, Inc. [References: 47]
机译:杆状病毒表达载体通常用于外源基因表达,并且作为改良的生物杀虫剂正在大力开发中。常规杆状病毒表达载体是重组病毒,其可以在多角体蛋白启动子的控制下在昆虫细胞中表达外源基因,该多角体蛋白启动子在感染的晚期提供高水平的转录。对于某些应用,包括外来糖蛋白生产和害虫防治,拥有可以在未感染细胞中或感染后更早表达外源基因产物的杆状病毒载体可能是有利的。为了满足这一需求,我们构建了一套新的质粒,可用于在杆状病毒iel启动子的控制下克隆和表达外源基因,该启动子在未感染的昆虫细胞和整个感染过程中均具有活性。我们使用这些新质粒的子集来分离含有各种外源基因的重组杆状病毒,并通过产生的即刻早期杆状病毒载体和常规杆状病毒载体比较了这些基因的表达。如预期的那样,即刻早期载体开始在感染中更早地表达每个外源基因,但是到感染后36-48小时,常规载体已经产生了更多的每种外源蛋白质。常规杆状病毒载体还比两个早期的杆状病毒载体从两个不同的原核基因产生了更多的酶活性。然而,即使在感染后48小时,即刻早期载体也能从两种不同的编码分泌途径蛋白的真核基因中产生比常规载体更多或更多的酶活性。因此,本报告描述了一套新的质粒,可用于在杆状病毒iel启动子的控制下克隆和表达外源基因,并建议立即早期的杆状病毒载体可能与常规杆状病毒表达载体一样有用,可用于生产具有生物活性的真核生物分泌途径蛋白。 (C)1996 Academic Press,Inc. [参考:47]

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