Identification of three extra-chromosomal replicons in Leptospira pathogenic strain and development of new shuttle vectors.
Identification of three extra-chromosomal replicons in Leptospira pathogenic strain and development of new shuttle vectors.
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钩端螺旋体致病菌株中三个染色体外复制子的鉴定和新穿梭载体的开发。
DOI：
10.1186/s12864-015-1321-y
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发表时间：
2015-02-15
期刊：
影响因子：
4.4
通讯作者：
Qin J
中科院分区：
文献类型：
--
作者：
Zhu W;Wang J;Zhu Y;Tang B;Zhang Y;He P;Zhang Y;Liu B;Guo X;Zhao G;Qin J
背景：致病性问号钩端螺旋体基因组含有两条染色体。已知质粒和原噬菌体在细菌的基因转移中发挥特定作用，并可能在这些生物体中作为有效的遗传工具。结果：在钩体中发现了3个染色体外复制子，分别命名为lcp 1（65，732 bp）、lcp 2（56，757 bp）和lcp 3（54，986 bp），它们的结构和功能在钩体遗传转化系统中的应用还没有得到充分的评价。通过全基因组测序鉴定了问号血清型临海株56609。所有三种复制子在细菌染色体外都是稳定的。在丝裂霉素C诱导后的56609培养上清中观察到噬菌体颗粒，并且含有噬菌体相关基因的lcp 3被认为是可诱导的前噬菌体。L.用来自三种质粒的单个rep或rep与parAB位点组合的预测复制元件构建的问号-大肠杆菌穿梭载体显示成功地转化到嗜血性和致病性钩端螺旋体物种中，表明rep基因在支持质粒的自动复制中的重要功能。此外，这三个rep基因的同源物在L.相关性检验表明，穿梭载体在问号线虫中的转化能力较好。结论：问号线虫分离株中同时存在3种染色体外复制子。其中一种我们认为是诱导型前噬菌体。用这三种复制子的rep基因构建的载体成功地转化到嗜血性和致病性钩端螺旋体物种中，但这部分取决于质粒和宿主的rep序列之间的遗传相容性。
Background:The genome of pathogenic Leptospira interrogans contains two chromosomes. Plasmids and prophages are known to play specific roles in gene transfer in bacteria and can potentially serve as efficient genetic tools in these organisms. Although plasmids and prophage remnants have recently been reported in Leptospira species, their characteristics and potential applications in leptospiral genetic transformation systems have not been fully evaluated.Results:Three extrachromosomal replicons designated lcp1 (65,732 bp), lcp2 (56,757 bp), and lcp3 (54,986 bp) in the L. interrogans serovar Linhai strain 56609 were identified through whole genome sequencing. All three replicons were stable outside of the bacterial chromosomes. Phage particles were observed in the culture supernatant of 56609 after mitomycin C induction, and lcp3, which contained phage-related genes, was considered to be an inducible prophage. L. interrogans-Escherichia coli shuttle vectors, constructed with the predicted replication elements of single rep or rep combined with parAB loci from the three plasmids were shown to successfully transform into both saprophytic and pathogenic Leptospira species, suggesting an essential function for rep genes in supporting auto-replication of the plasmids. Additionally, a wide distribution of homologs of the three rep genes was identified in L. interrogans isolates, and correlation tests showed that the transformability of the shuttle vectors in L. interrogans isolates depended, to certain extent, on genetic compatibility between the rep sequences of both plasmid and host.Conclusions:Three extrachromosomal replicons co-exist in L. interrogans, one of which we consider to be an inducible prophage. The vectors constructed with the rep genes of the three replicons successfully transformed into saprophytic and pathogenic Leptospira species alike, but this was partly dependent on genetic compatibility between the rep sequences of both plasmid and host.
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影响因子：
3.2
作者：
Humphrey, SB;Stanton, TB;Zuerner, RL
通讯作者：
Zuerner, RL
影响因子：
14.9
作者：
Finn RD;Mistry J;Tate J;Coggill P;Heger A;Pollington JE;Gavin OL;Gunasekaran P;Ceric G;Forslund K;Holm L;Sonnhammer EL;Eddy SR;Bateman A
通讯作者：
Bateman A
影响因子：
4.2
作者：
Liao S;Sun A;Ojcius DM;Wu S;Zhao J;Yan J
通讯作者：
Yan J
影响因子：
3.1
作者：
Bourhy, Pascale;Salaun, Laurence;Picardeau, Mathieu
通讯作者：
Picardeau, Mathieu
影响因子：
3.7
作者：
Canchaya, C;Desiere, F;Brüssow, H
通讯作者：
Brüssow, H