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首页> 外文期刊>Applied Microbiology >Involvement of the Major Capsid Protein and Two Early-Expressed Phage Genes in the Activity of the Lactococcal Abortive Infection Mechanism AbiT
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Involvement of the Major Capsid Protein and Two Early-Expressed Phage Genes in the Activity of the Lactococcal Abortive Infection Mechanism AbiT

机译：主要衣壳蛋白和两个早期表达的噬菌体基因参与乳球菌流产感染机制AbiT的活动。

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The dairy industry uses the mesophilic, Gram-positive, lactic acid bacterium (LAB) Lactococcus lactis to produce an array of fermented milk products. Milk fermentation processes are susceptible to contamination by virulent phages, but a plethora of phage control strategies are available. One of the most efficient is to use LAB strains carrying phage resistance systems such as abortive infection (Abi) mechanisms. Yet, the mode of action of most Abi systems remains poorly documented. Here, we shed further light on the antiviral activity of the lactococcal AbiT system. Twenty-eight AbiT-resistant phage mutants derived from the wild-type AbiT-sensitive lactococcal phages p2, bIL170, and P008 were isolated and characterized. Comparative genomic analyses identified three different genes that were mutated in these virulent AbiT-insensitive phage derivatives: e14 (bIL170 [ e14 _(bIL170)]), orf41 (P008 [ orf41 _(P008)]), and orf6 (p2 [ orf6 _(p2)] and P008 [ orf6 _(P008)]). The genes e14 _(bIL170) and orf41 _(P008) are part of the early-expressed genomic region, but bioinformatic analyses did not identify their putative function. orf6 is found in the phage morphogenesis module. Antibodies were raised against purified recombinant ORF6, and immunoelectron microscopy revealed that it is the major capsid protein (MCP). Coexpression in L. lactis of ORF6_(p2) and ORF5_(p2), a protease, led to the formation of procapsids. To our knowledge, AbiT is the first Abi system involving distinct phage genes.
机译：乳制品行业使用嗜温，革兰氏阳性的乳酸菌（LAB）乳酸乳球菌生产一系列发酵的乳制品。牛奶发酵过程容易受到有毒噬菌体的污染，但是可以使用多种噬菌体控制策略。最有效的方法之一是使用带有噬菌体抗性系统的LAB菌株，例如流产感染（Abi）机制。但是，大多数Abi系统的作用方式仍然记录不清。在这里，我们进一步揭示了乳球菌AbiT系统的抗病毒活性。分离并鉴定了来自野生型AbiT敏感的乳球菌噬菌体p2，bIL170和P008的28个抗AbiT的噬菌体突变体。比较基因组分析确定了在这些高毒力的AbiT不敏感噬菌体衍生物中突变的三个不同基因：e14（bIL170 [e14 _（bIL170）]），orf41（P008 [orf41 _（P008）]）和orf6（p2 [orf6 _） （p2）]和P008 [orf6 _（P008）]）。基因e14 _（bIL170）和orf41 _（P008）是早期表达的基因组区域的一部分，但生物信息学分析未能确定其推定功能。在噬菌体形态发生模块中发现了orf6。产生了针对纯化的重组ORF6的抗体，免疫电子显微镜显示这是主要的衣壳蛋白（MCP）。乳酸乳球菌中的蛋白酶ORF6_（p2）和ORF5_（p2）共表达导致了衣壳的形成。据我们所知，AbiT是第一个涉及独特噬菌体基因的Abi系统。

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