Multiple large foreign protein expression by a single recombinant baculovirus: A system for production of multivalent vaccines
Multiple large foreign protein expression by a single recombinant baculovirus: A system for production of multivalent vaccines
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DOI：
10.1016/j.pep.2013.07.005
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发表时间：
2013-09-01
影响因子：
1.6
通讯作者：
Roy, Polly
中科院分区：
文献类型：
--
作者：
Kanai, Yuta;Athmaram, T. N.;Roy, Polly
杆状病毒表达系统提供了通过单个重组病毒同时表达几种大蛋白的优点。迄今为止，由于需要产生大的构建体和重复使用同源序列和启动子，多种大（>100 kDa）蛋白质的表达受到阻碍。多位点杆状病毒表达系统的发展通过使大的外源序列能够重组到基因组的不同区域来克服这些问题。在本文中，我们研究了非洲马瘟病毒（AHSV）VP2蛋白的共表达从多个血清型在一个单一的重组杆状病毒。为此，产生了表达多种AHSV VP2蛋白的重组杆状病毒，并且发现可以从杆状病毒基因组的不同基因座同时表达多达六种不同的AHSV血清型（血清型1、3、4、5、7和8）VP2蛋白（类似于120kDa）。一种血清型的VP2的表达并没有受到其他血清型的存在的显著阻碍，尽管不同血清型之间的表达水平略有差异。来自其他基因座的其他血清型的VP2的表达导致VP2蛋白的表达水平降低。基于这些发现，构建了包含所有9种AHSV血清型（血清型1、7、8或血清型2、4、5或血清型3、6、9）的另外3种重组杆状病毒，并且每种三重重组病毒均显示出每种VP2的相似表达水平。该系统允许表达许多大的蛋白质，其具有用于多价疫苗生产的潜力。(C)2013 Elsevier Inc. All rights reserved.
Baculovirus expression system offers the advantage of expression of several large proteins simultaneously by a single recombinant virus. To date, expression of multiple large (>100 kDa) proteins has been hampered by the need to generate large constructs and repeat use of homologous sequence and promoter. The development of multi-loci baculovirus expression system overcomes these issues by enabling the recombination of large foreign sequences into different regions of the genome. In this paper, we have examined the co-expression of African horse sickness virus (AHSV) VP2 proteins from multiple serotypes in a single recombinant baculovirus. To this end, recombinant baculoviruses expressing multiple AHSV VP2 proteins were generated and it was found that up to six different AHSV serotypes (serotype 1, 3, 4, 5, 7 and 8) VP2 proteins (similar to 120 kDa) could be expressed simultaneously from different loci of baculovirus genome. The expression of VP2 of one serotype was not significantly hindered by the presence of other serotypes, although there were slight differences in expression level between different serotypes. The expression of VP2 of further serotypes from additional loci resulted in a lesser expression level of VP2 proteins. Based on these findings, three additional recombinant baculoviruses encompassing all nine AHSV serotypes were constructed (serotypes 1, 7, 8 or serotypes 2, 4, 5 or serotypes 3, 6, 9) and each of the triple recombinant viruses exhibited similar expression level of each VP2. This system allows for the expression of a number of large proteins that has the potential to be exploited for multivalent vaccines production. (C) 2013 Elsevier Inc. All rights reserved.