Bieńkowska-Szewczyk K, Szewczyk B
Department of Molecular Virology, Faculty of Biotechnology, University of Gdańsk, Poland.
Acta Biochim Pol. 1999;46(2):325-39.
The outermost layers of animal viruses are usually composed of glycoproteins. They are responsible not only for the entrance of viruses into, and release from host cells but also for the initial interaction of a viral particle with immunological defense of the host. It is therefore not surprising that many laboratories devote a lot of effort to study viral glycoproteins at the molecular level. Very often such studies are possible only after the introduction of a glycoprotein gene into a heterologous system. Expression of glycoprotein genes is usually obtained in mammalian or insect cells. Expression in mammalian cells yields viral glycoproteins with glycan chains indistinguishable from the original counterparts in virion particles but the level of synthesis of glycoproteins is very low. Vaccinia virus is the most common vector for expression in mammalian cells. It is easy to grow, the introduction of foreign genes is relatively simple and, due to the size of the vaccinia genome, it can accept large pieces of foreign DNA. Glycosylation in insect cells is not as complex as in mammalian cells and usually glycoproteins produced in insect cells are of slightly lower molecular mass than those produced in mammalian cells. The most common vector for expression of glycoproteins in insect cells is a baculovirus, Autographa californica nuclear polyhedrosis virus (AcNPV). The great advantage of this system is a very high level of expression of foreign genes.
动物病毒的最外层通常由糖蛋白组成。它们不仅负责病毒进入宿主细胞以及从宿主细胞中释放,还负责病毒颗粒与宿主免疫防御的初始相互作用。因此,许多实验室致力于在分子水平上研究病毒糖蛋白也就不足为奇了。通常只有在将糖蛋白基因导入异源系统后,此类研究才有可能进行。糖蛋白基因的表达通常在哺乳动物细胞或昆虫细胞中实现。在哺乳动物细胞中表达可产生具有与病毒体颗粒中原始对应物难以区分的糖链的病毒糖蛋白,但糖蛋白的合成水平非常低。痘苗病毒是在哺乳动物细胞中表达最常用的载体。它易于培养,导入外源基因相对简单,并且由于痘苗病毒基因组的大小,它可以接受大片段的外源DNA。昆虫细胞中的糖基化不像哺乳动物细胞中那么复杂,通常昆虫细胞中产生的糖蛋白分子量略低于哺乳动物细胞中产生的糖蛋白。在昆虫细胞中表达糖蛋白最常用的载体是杆状病毒,苜蓿银纹夜蛾核型多角体病毒(AcNPV)。该系统的最大优势是外源基因的表达水平非常高。