Identification of the genes involved in Riemerella anatipestifer biofilm formation by random transposon mutagenesis.
Identification of the genes involved in Riemerella anatipestifer biofilm formation by random transposon mutagenesis.
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通过随机转座子诱变鉴定参与鸭疫里默氏菌生物膜形成的基因
DOI：
10.1371/journal.pone.0039805
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Yu S
中科院分区：
文献类型：
--
作者：
Hu Q;Zhu Y;Tu J;Yin Y;Wang X;Han X;Ding C;Zhang B;Yu S
鸭疫里默氏杆菌是一种引起家禽流行的传染病，给养鸭业造成严重的经济损失。我们以前的研究表明，一些菌株的R。鸭疫杆菌可以形成生物膜，这可能解释了R.感染后鸭场的鸭疫病毒。本研究以生物被膜产生菌CH 3为材料，构建了转座子Tn 4351随机突变体库，以研究生物被膜形成的遗传基础。在非生物表面上的寄生虫。共获得2，520个突变体，其中39个使用结晶紫染色显示生物膜形成减少47%-98%。突变体的遗传特性导致33个基因的鉴定。其中，29个基因与信息存储和处理以及基本的细胞过程和代谢有关;其他四个基因的功能目前尚不清楚。此外，用穿梭质粒pCP-dhdps补充突变菌株BF 19，其中在二氢吡啶二羧酸合酶（dhdps）基因处插入Tn 4351转座子后生物膜形成减少98%。互补突变株恢复到野生型菌株CH 3的92.6%的生物膜形成，这表明dhdp基因与生物膜形成相关。据推断，这种互补作用也适用于其他突变株。此外，对生物膜缺陷突变株的生物学特性进行了研究，发现突变株中缺失的基因在生物膜形成过程中起作用。鸭疫任一基因的缺失将在特定阶段停止生物膜形成，从而防止进一步的生物膜发展。此外，测试的生物膜缺陷型突变体对Vero细胞的粘附能力不同。本研究将有助于我们了解红曲霉生物膜形成的分子机制。研究鸭疫杆菌的致病机理。anatipestifer进一步。
Riemerella anatipestifer causes epizootics of infectious disease in poultry that result in serious economic losses to the duck industry. Our previous studies have shown that some strains of R. anatipestifer can form a biofilm, and this may explain the intriguing persistence of R. anatipestifer on duck farms post infection. In this study we used strain CH3, a strong producer of biofilm, to construct a library of random Tn4351 transposon mutants in order to investigate the genetic basis of biofilm formation by R. anatipestifer on abiotic surfaces. A total of 2,520 mutants were obtained and 39 of them showed a reduction in biofilm formation of 47%–98% using crystal violet staining. Genetic characterization of the mutants led to the identification of 33 genes. Of these, 29 genes are associated with information storage and processing, as well as basic cellular processes and metabolism; the function of the other four genes is currently unknown. In addition, a mutant strain BF19, in which biofilm formation was reduced by 98% following insertion of the Tn4351 transposon at the dihydrodipicolinate synthase (dhdps) gene, was complemented with a shuttle plasmid pCP-dhdps. The complemented mutant strain was restored to give 92.6% of the biofilm formation of the wild-type strain CH3, which indicates that the dhdp gene is associated with biofilm formation. It is inferred that such complementation applies also to other mutant strains. Furthermore, some biological characteristics of biofilm-defective mutants were investigated, indicating that the genes deleted in the mutant strains function in the biofilm formation of R. anatipestifer. Deletion of either gene will stall the biofilm formation at a specific stage thus preventing further biofilm development. In addition, the tested biofilm-defective mutants had different adherence capacity to Vero cells. This study will help us to understand the molecular mechanisms of biofilm development by R. anatipestifer and to study the pathogenesis of R. anatipestifer further.
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DOI：
10.1038/nbt1183-784
发表时间：
1983-01-01
期刊：
BIO-TECHNOLOGY
影响因子：
--
作者：
SIMON, R;PRIEFER, U;PUHLER, A
通讯作者：
PUHLER, A
影响因子：
3.2
作者：
Cooper, AJ;Kalinowski, AP;Salyers, AA
通讯作者：
Salyers, AA
DOI：
10.1073/pnas.86.7.2172
发表时间：
1989-04-01
影响因子：
11.1
作者：
CHUNG, CT;NIEMELA, SL;MILLER, RH
通讯作者：
MILLER, RH
影响因子：
4.4
作者：
Alvarez, B.;Secades, P.;Guijarro, J. A.
通讯作者：
Guijarro, J. A.
影响因子：
3.1
作者：
Cuccui, J.;Easton, A.;Wren, B. W.
通讯作者：
Wren, B. W.