Heterologous expression of Shigella dysenteriae serotype 1 O-antigen analog in Escherichia coli K-12 W3110 by transferring a minimum number of genes involved in O-polysaccharide biosynthesis
Heterologous expression of Shigella dysenteriae serotype 1 O-antigen analog in Escherichia coli K-12 W3110 by transferring a minimum number of genes involved in O-polysaccharide biosynthesis
复制标题
通过转移参与 O-多糖生物合成的最小数量的基因，在大肠杆菌 K-12 W3110 中异源表达志贺氏菌血清型 1 O-抗原类似物
DOI：
10.1007/s10529-018-2584-6
复制
发表时间：
2018-06
影响因子：
2.7
通讯作者：
Chen Min
中科院分区：
文献类型：
--
作者：
Kong Yun;Qu Yajun;Wang Shengjun;Wang Peng George;Chen Min
目的：用最少数量的基因代替整个O-PS基因簇，在大肠杆菌中异源产生痢疾志贺氏菌血清型1 o -多糖（O-PS， o -抗原）。结果：将负责o重复单元形成的3个糖基转移酶基因（rfbR、rfbQ和rfp）导入大肠杆菌K-12 W3110中，合成了S. dysenteriae 1 O-PS。利用SDS-PAGE银染色和S. dysenteriae 1脂多糖抗血清免疫印迹法，在重组大肠杆菌中观察到具有不同o重复单元链长的特异性o抗原阶梯型。质谱和离子色谱分析表明，产生了带有额外葡萄糖残基的特定S. dysenteriae 1 O-repeat单元结构。结论：表达特异性糖基转移酶基因的重组大肠杆菌可产生痢疾链球菌1型O-PS，并可合成多种致病菌的异源o -抗原用于疫苗制备。
Objective:To heterologously produce the Shigella dysenteriae serotype 1 O-polysaccharide (O-PS, O-antigen) in Escherichia coli by transferring the minimum number of genes instead of the entire O-PS gene cluster.Results:The three glycosyltransferase genes (rfbR, rfbQ and rfp) responsible for the formation of the O-repeat unit were introduced into E. coli K-12 W3110 to synthesize S. dysenteriae 1 O-PS. The specific O-antigen ladder type with different chain lengths of O-repeat units was observed in the recombinant E. coli strain by SDS-PAGE silver staining and western blotting using S. dysenteriae 1 lipopolysaccharide antiserum. Analysis by mass spectrometry and ion chromatography suggested generation of the specific S. dysenteriae 1 O-repeat unit structure with an extra glucose residue attached.Conclusions:Recombinant E. coli expressing specific glycosyltransferase genes can generate the O-PS of S. dysenteriae 1 and might be able to synthesize heterologous O-antigens of various pathogenic bacteria for vaccine preparation.
登录
查看更多内容
影响因子：
5.5
作者：
De-qi Xu;J. Cisar;M. Osorio;T. Wai;D. Kopecko
通讯作者：
De-qi Xu;J. Cisar;M. Osorio;T. Wai;D. Kopecko
影响因子：
3.2
作者：
KLENA, JD;ASHFORD, RS;SCHNAITMAN, CA
通讯作者：
SCHNAITMAN, CA
影响因子：
4.4
作者：
Ilkka Seppälä;O. Mäkelä
通讯作者：
Ilkka Seppälä;O. Mäkelä
影响因子：
3.1
作者：
PEETERS, CCAM;EVENBERG, D;POOLMAN, JT
通讯作者：
POOLMAN, JT
影响因子：
4.3
作者：
Ravenscroft, Neil;Haeuptle, Micha A.;Wacker, Michael
通讯作者：
Wacker, Michael