Sequence and characterization of shuttle vectors for molecular cloning in Porphyromonas, Bacteroides and related bacteria.
Sequence and characterization of shuttle vectors for molecular cloning in Porphyromonas, Bacteroides and related bacteria.
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DOI：
10.1111/omi.12304
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发表时间：
2020-08
影响因子：
3.7
通讯作者：
Lewis JP
中科院分区：
文献类型：
--
作者：
Jones KR;Belvin BR;Macrina FL;Lewis JP
牙龈卟啉单胞菌和相关物种的遗传学研究所需的穿梭载体缺乏。为了更好地了解这些工具的候选者的流行情况，我们检查了多株黑色厌氧菌（临床和实验室分离株）的质粒。由于在牙龈卟啉单胞菌菌株中没有发现质粒，我们使用来自解砷卟啉单胞菌的pYH 420质粒作为骨架与来自大肠杆菌的pUC 19组合构建穿梭载体。pYH 420质粒的核苷酸序列测定显示，它含有与来自质粒pTS 1（分离自齿垢密螺旋体）的rep相似的基因以及mobA的同源物，mobA是在拟杆菌属中发现的可移动遗传元件上发现的基因家族的成员。我们利用pUC 19和pYH 420载体的一部分构建了pG 106和pG 108穿梭载体。这导致了在lacZ基因中具有多克隆位点（MCS）的载体，使我们能够进行蓝白菌落选择。pG 106和pG 108穿梭载体可电转化到E.大肠杆菌、牙龈卟啉单胞菌和B. thetaiotaomicron，在那里他们是稳定的。我们证明，这些载体是适合在这些物种的分子克隆，包括互补和基因表达研究的应用。使用pG 108载体，我们补充了牙龈卟啉单胞菌的hcpR突变株，并挽救了其敏感表型。我们还在B中使用P-glow BS 2荧光报告基因和ahpC启动子进行了基因表达研究。太多了。
There is a lack of shuttle vectors to be needed for investigations into the genetics of Porphyromonas gingivalis and related species. To better understand the prevalence of candidates for such tools, we have examined multiple strains of black-pigmented anaerobes (clinical and laboratory isolates) for plasmids. As no plasmids were found in P. gingivalis strains, we have used the pYH420 plasmid, derived from P. asaccharolytica, as backbone to construct a shuttle vector in combination with pUC19 from Escherichia coli. Nucleotide sequence determination of the pYH420 plasmid revealed that it contained a gene with similarity to rep from plasmid pTS1 (isolated from Treponema denticola) as well as a homolog of mobA, a member of a gene family found on mobilizable genetic elements found in the genus Bacteroides. We constructed the pG106 and pG108 shuttle vectors using parts of the pUC19 and pYH420 vectors. This resulted in a vector with a multiple cloning site (MCS) in the lacZ gene enabling us to perform blue–white colony selection. The pG106 and pG108 shuttle vectors are electro-transformable into E. coli, P. gingivalis and B. thetaiotaomicron, where they are stable. We demonstrated that these vectors were suitable in these species for applications of molecular cloning including complementation and gene expression studies. Using the pG108 vector, we complement the hcpR mutant strain of P. gingivalis and rescued its -sensitive phenotype. We also performed a gene expression study using the P-glow BS2 fluorescent reporter gene and the ahpC promoter in B. thetaiotaomicron.
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Keystone-Pathogen假设。
DOI：
10.1038/nrmicro2873
发表时间：
2012-10
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
通讯作者：
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影响因子：
5.2
作者：
How KY;Song KP;Chan KG
通讯作者：
Chan KG
影响因子：
4.4
作者：
Gardner, RG;Russell, JB;Shoemaker, NB
通讯作者：
Shoemaker, NB
DOI：
10.1016/j.tripleo.2006.05.010
发表时间：
2007-02-01
期刊：
ORAL SURGERY ORAL MEDICINE ORAL PATHOLOGY ORAL RADIOLOGY AND ENDODONTOLOGY
影响因子：
--
作者：
Tomazinho, Luiz Fernando;Avila-Campos, Mario J.
通讯作者：
Avila-Campos, Mario J.
影响因子：
3.1
作者：
Chen, TT;Dong, H;Duncan, MJ
通讯作者：
Duncan, MJ