CLONING AND MOLECULAR ANALYSIS OF THE GALE GENE OF NEISSERIA-MENINGITIDIS AND ITS ROLE IN LIPOPOLYSACCHARIDE BIOSYNTHESIS
CLONING AND MOLECULAR ANALYSIS OF THE GALE GENE OF NEISSERIA-MENINGITIDIS AND ITS ROLE IN LIPOPOLYSACCHARIDE BIOSYNTHESIS
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DOI：
10.1111/j.1749-6632.1994.tb44256.x
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发表时间：
1994-01-01
期刊：
影响因子：
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通讯作者：
MOXON, ER
中科院分区：
文献类型：
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作者：
JENNINGS, MP;VANDERLEY, P;MOXON, ER
脑膜炎双球菌脑膜炎和败血症仍然是世界范围内的重大健康问题。现有的疫苗对A组和C组菌株的保护作用有限，但对B组生物没有保护作用，而B组生物是许多国家疾病的主要原因。脑膜炎奈瑟氏菌的脂多糖(LPS)被认为是毒力和宿主抵抗感染的主要决定因素。*在LPS中存在含有半乳糖的结构，Gala I-4galp和lacto-N-neotetraose，它们也存在于上皮细胞表面或宿主分泌物中。3A这些结构在脑膜炎奈瑟菌感染发病机制中的生物学作用尚不清楚。本研究以伊氏嗜血杆菌的gule基因作为B群脑膜炎奈瑟菌的gule基因的杂交探针，鉴定了两个不同的同源基因座。对每个基因座进行了克隆，核苷酸序列分析表明，两个基因座都含有与GUE相似的序列。其中一个含有功能基因gule，定位于被膜生物合成位点。第二个片段只包含部分GULE编码序列，不表达功能基因产物。通过转化灭活的gule基因构建了一株gule突变型脑膜炎双球菌。对来自gule突变株的内毒素的分析显示，分子量明显减少(图1)，并与已知含有半乳糖的L3和L8免疫类型的单抗失去反应能力。这些结果与脂多糖中半乳糖的损失是一致的，并定义了半乳糖-
Meningococcal meningitis and septicemia remain significant health problems worldwide. Vaccines are available which give a limited degree of protection against Group A and C strains, but not the Group B organisms, which are the major cause of disease in many countries. The lipopolysaccharide (LPS) of Neisseriu meningitidis is known to be a major determinant of virulence'and of host defense against infection.* Present in the LPS are the galactose-containing structures, gala I-4galp and lacto-N-neotetraose, which are also found on the surface of epithelial cells or in host secretions. 3A The biologic role of these structures in the pathogenesis of N. meningitidis infection is not known.In this study, the gulE gene from Huemophilus injluenzue was used as a hybridization probe for the gulE gene of N. meningitidis Group B, identifying two different homologous loci. Each of the loci was cloned, and nucleotide sequence analysis revealed that both loci contained sequences similar to gulE. One contained a functional gulE gene and mapped to the capsule biosynthetic locus. The second contained only a partial gulE coding sequence which did not express a functional gene product. A gulE mutant meningococcal strain was constructed by transformation with an inactivated gulE gene. Analysis of the LPS from the gulE mutant strain revealed an apparent reduction in molecular weight (FIG. 1) and a loss of reactivity with monoclonal antibodies specific for L3 and L8 immunotypes known to contain galactose. These results are consistent with the loss of galactose from the LPS and defines the galactose-