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Conjugative Plasmid from Lactobacillus gasseri LA39 That Carries Genes for Production of and Immunity to the Circular Bacteriocin Gassericin A

机译:Gasseri LA39乳酸杆菌的结合质粒携带环状细菌Gastroericin A的生产和免疫基因

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摘要

Gassericin A is a circular bacteriocin produced by Lactobacillus gasseri strain LA39. We found a 33,333-bp plasmid, designated pLgLA39, in this strain. pLgLA39 contained 44 open reading frames, including seven genes related to gassericin A production/immunity (gaa), as well as genes for replication, plasmid maintenance, and conjugative transfer. pLgLA39 was transferred from LA39 to the type strain of L. gasseri (JCM 1131) by filter mating. The transconjugant exhibited >30-fold-higher more resistance to gassericin A and produced antibacterial activity. Lactobacillus reuteri LA6, the producer of reutericin 6, was proved to harbor a plasmid indistinguishable from pLgLA39 and carrying seven genes 100% identical to gaa. This suggests that pLgLA39 might have been transferred naturally between L. gasseri LA39 and L. reuteri LA6. The seven gaa genes of pLgLA39 were cloned into a plasmid vector to construct pGAA. JCM 1131T transformed with pGAA expressed antibacterial activity and resistance to gassericin A. pGAA was segregationally more stable than a pGAA derivative plasmid from which gaaA was deleted and even was more stable than the vector. This suggests the occurrence of postsegregational host killing by the gaa genes. pLgLA39 carried a pemIK homolog, and segregational stabilization of a plasmid by the pLgLA39-type pemIK genes was also confirmed. Thus, pLgLA39 was proved to carry the genes for at least two plasmid maintenance mechanisms, i.e., gaa and pemIK. Plasmids containing a repA gene similar to pLgLA39 repA were distributed in several L. gasseri strains.
机译:Gassericin A是由Gasseri gasseri菌株LA39产生的环状细菌素。我们在该菌株中发现了一个33333 bp的质粒,命名为pLgLA39。 pLgLA39包含44个开放阅读框,包括七个与Gassericin A产生/免疫(gaa)相关的基因,以及用于复制,质粒维持和结合转移的基因。通过过滤器配合将pLgLA39从LA39转移到加氏乳杆菌的菌株(JCM 1131)。该转导结合剂对加色霉素A的抗性高出> 30倍,并具有抗菌活性。已证明罗伊氏乳杆菌6的生产者罗伊氏乳杆菌LA6含有与pLgLA39不能区分的质粒,并携带7个与gaa 100%相同的基因。这表明pLgLA39可能已经在加氏乳杆菌LA39和罗伊氏乳杆菌LA6之间自然转移。将pLgLA39的七个gaa基因克隆到质粒载体中以构建pGAA。用pGAA转化的JCM 1131 T 表现出抗菌活性和对Gassericin A的抗性。pGAA在分离性上比缺失gaaA的pGAA衍生质粒更稳定,甚至比载体更稳定。这表明分离后宿主被gaa基因杀死的现象。 pLgLA39带有pemIK同源物,并且还证实了pLgLA39型pemIK基因对质粒的分离稳定作用。因此,证明pLgLA39携带至少两个维持质粒机制的基因,即gaa和pemIK。含有类似于pLgLA39 repA的repA基因的质粒分布在几个 L中。 Gasseri 菌株。

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