Overexpression of Brucella putative glycosyltransferase WbkA in B. abortus RB51 leads to production of exopolysaccharide.
Overexpression of Brucella putative glycosyltransferase WbkA in B. abortus RB51 leads to production of exopolysaccharide.
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DOI：
10.3389/fcimb.2015.00054
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发表时间：
2015
影响因子：
5.7
通讯作者：
Vemulapalli R
中科院分区：
文献类型：
--
作者：
Dabral N;Jain-Gupta N;Seleem MN;Sriranganathan N;Vemulapalli R
布鲁氏菌是革兰氏阴性、兼性胞内细菌，在哺乳动物中引起布鲁氏菌病。在其细胞壁结构中含有O-多糖的布鲁氏菌菌株表现出光滑的表型，而缺乏多糖的菌株表现出粗糙的表型。B。牛种布氏杆菌RB 51株是一株稳定的粗毒弱毒突变株，可作为获得许可的牛种布氏杆菌病活疫苗使用。先前的研究表明，编码O-多糖合成所需的糖基转移酶的wboA基因在B中被破坏。流产RB 51的IS 711元件。尽管菌株RB 51与功能性wboA基因的互补导致细胞质中的O-多糖合成，但其不导致平滑表型。本研究的目的是确定是否过表达布鲁氏菌WbkA或WbkE，两个额外的推定的糖基转移酶的O-多糖合成所必需的，在菌株RB 51将导致O-多糖合成和光滑的表型。我们的研究结果表明，RB 51中wbkA或wbkE基因的过表达不会导致O-多糖的表达，如用特异性抗体进行的Western印迹所示。然而，wbkA而不是wbkE的过表达导致凝集表型的发展和含有甘露糖、半乳糖、N-乙酰葡糖胺和N-乙酰半乳糖胺的胞外多糖的产生。此外，我们发现凝集重组菌株显示出对聚苯乙烯板的粘附增加。重组菌株在减毒特性和诱导抗强毒B保护性免疫的能力方面与菌株RB 51相似。小鼠流产攻毒。
Brucella spp. are Gram-negative, facultative intracellular bacteria that cause brucellosis in mammals. Brucella strains containing the O-polysaccharide in their cell wall structure exhibit a smooth phenotype whereas the strains devoid of the polysaccharide show rough phenotype. B. abortus strain RB51 is a stable rough attenuated mutant which is used as a licensed live vaccine for bovine brucellosis. Previous studies have shown that the wboA gene, which encodes a glycosyltransferase required for the synthesis of O-polysaccharide, is disrupted in B. abortus RB51 by an IS711 element. Although complementation of strain RB51 with a functional wboA gene results in O-polysaccharide synthesis in the cytoplasm, it does not result in smooth phenotype. The aim of this study was to determine if overexpression of Brucella WbkA or WbkE, two additional putative glycosyltransferases essential for O-polysaccharide synthesis, in strain RB51 would result in the O-polysaccharide synthesis and smooth phenotype. Our results demonstrate that overexpression of wbkA or wbkE gene in RB51 does not result in O-polysaccharide expression as shown by Western blotting with specific antibodies. However, wbkA, but not wbkE, overexpression leads to the development of a clumping phenotype and the production of exopolysaccharide(s) containing mannose, galactose, N-acetylglucosamine, and N-acetylgalactosamine. Moreover, we found that the clumping recombinant strain displays increased adhesion to polystyrene plates. The recombinant strain was similar to strain RB51 in its attenuation characteristic and in its ability to induce protective immunity against virulent B. abortus challenge in mice.
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影响因子：
3.1
作者：
McQuiston, JR;Vemulapalli, R;Boyle, SM
通讯作者：
Boyle, SM
影响因子：
3.7
作者：
González D;Grilló MJ;De Miguel MJ;Ali T;Arce-Gorvel V;Delrue RM;Conde-Alvarez R;Muñoz P;López-Goñi I;Iriarte M;Marín CM;Weintraub A;Widmalm G;Zygmunt M;Letesson JJ;Gorvel JP;Blasco JM;Moriyón I
通讯作者：
Moriyón I
DOI：
10.2174/1874285801307010087
发表时间：
2013
期刊：
The open microbiology journal
影响因子：
--
作者：
Almirón MA;Roset MS;Sanjuan N
通讯作者：
Sanjuan N
影响因子：
3.1
作者：
CAROFF, M;BUNDLE, DR;DUNCAN, JR
通讯作者：
DUNCAN, JR
影响因子：
3.7
作者：
Adone, Rosanna;Muscillo, Michele;Tarantino, Michela
通讯作者：
Tarantino, Michela