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首页> 外文期刊>Infection and immunity >Expression of a cloned lipopolysaccharide antigen from Neisseria gonorrhoeae on the surface of Escherichia coli K-12.
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Expression of a cloned lipopolysaccharide antigen from Neisseria gonorrhoeae on the surface of Escherichia coli K-12.

机译：淋病奈瑟氏球菌克隆的脂多糖抗原在大肠杆菌K-12表面的表达。

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A gonococcal gene bank maintained in Escherichia coli K-12 was screened by colony immunoblotting, and a transformant expressing a surface antigen reactive to anti-gonococcal outer membrane antiserum was isolated. The isolate carried a recombinant plasmid, pTME6, consisting of approximately 9 kilobases of Neisseria gonorrhoeae DNA inserted into the BamHI site of pBR322. Surface labeling of E. coli HB101(pTME6) confirmed that the antigen was expressed on the E. coli cell surface. The antigenic material was resistant to proteinase K digestion and sensitive to periodate oxidation, indicating that the material was carbohydrate. Purified lipopolysaccharide (LPS) from HB101(pTME6) produced a unique band on silver-stained polyacrylamide gels that contained immunoreactive material as seen on Western blots of LPS samples. Only two of three E. coli LPS mutant strains carrying pTME6 reacted with the antigonococcal antiserum, suggesting that a certain E. coli core structure is necessary for antigen expression. We conclude that pTME6 contains one or more gonococcal genes encoding an LPS core biosynthetic enzyme(s) which can modify E. coli core LPS to produce a gonococcuslike epitope(s).
机译：通过菌落免疫印迹法筛选在大肠杆菌K-12中维持的淋球菌基因库，并分离出表达与抗淋球菌外膜抗血清反应的表面抗原的转化体。该分离物带有重组质粒pTME6，该质粒由插入pBR322的BamHI位点的约9千株淋病奈瑟氏球菌DNA组成。大肠杆菌HB101（pTME6）的表面标记证实该抗原在大肠杆菌细胞表面表达。抗原物质对蛋白酶K的消化有抵抗力，并对高碘酸盐氧化敏感，表明该物质是碳水化合物。纯化自HB101（pTME6）的脂多糖（LPS）在银染聚丙烯酰胺凝胶上产生了一条独特的条带，该条带含有免疫反应性物质，如LPS样品的蛋白质印迹所示。携带pTME6的3个大肠杆菌LPS突变菌株中只有2个与抗淋球菌抗血清反应，这表明一定的大肠杆菌核心结构对于抗原表达是必需的。我们得出的结论是，pTME6包含一种或多种编码LPS核心生物合成酶的淋球菌基因，该酶可以修饰大肠杆菌核心LPS产生淋球菌样抗原决定簇。

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