首页> 美国卫生研究院文献>Infection and Immunity >Cloning and characterization of a Chlamydia trachomatis L3 DNA fragment that codes for an antigenic region of the major outer membrane protein and specifically hybridizes to the C- and C-related-complex serovars.
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Cloning and characterization of a Chlamydia trachomatis L3 DNA fragment that codes for an antigenic region of the major outer membrane protein and specifically hybridizes to the C- and C-related-complex serovars.

机译:沙眼衣原体L3 DNA片段的克隆和表征该片段编码主要外膜蛋白的抗原区并与C和C相关的复杂血清型特异性杂交。

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摘要

Chlamydia trachomatis L3 DNA was cloned and expressed in lambda gt11. A recombinant plaque that expressed an antigen that reacted with rabbit polyclonal antichlamydial L3 serum and with two monoclonal antibodies specific for serovars L3 and I was selected from this Chlamydia genomic library. The beta-galactosidase Chlamydia fusion protein was purified by immunoaffinity chromatography and injected into mice to produce monoclonal antibodies. These monoclonal antibodies reacted by Western (immuno-) blot with both the fusion protein and the major outer membrane protein from purified L3 elementary bodies. The chlamydial DNA fragment was shown by DNA sequence analysis to be 168 base pairs in length and to correspond to the constant regions 1 and 2 and the variable segment 1 of the major outer membrane protein gene. The recombinant chlamydial DNA fragment hybridized under stringent conditions by Southern and dot blot analysis exclusively with the DNA from the C- and C-related-complex C. trachomatis serovars.
机译:克隆了沙眼衣原体L3 DNA,并在λgt11中表达。从该衣原体基因组文库中选择了表达抗原的重组噬菌斑,该抗原与兔多克隆抗衣原体L3血清和两种对血清型L3和I具有特异性的单克隆抗体反应。通过免疫亲和层析纯化β-半乳糖苷酶衣原体融合蛋白,并将其注射到小鼠中以产生单克隆抗体。这些单克隆抗体通过蛋白质印迹(免疫印迹)与来自纯化的L3基本体的融合蛋白和主要外膜蛋白反应。通过DNA序列分析显示衣原体DNA片段的长度为168个碱基对,并对应于主要外膜蛋白基因的恒定区1和2以及可变区1。重组衣原体DNA片段在严格条件下通过Southern和斑点印迹分析仅与来自C和C相关复杂沙眼衣原体血清型的DNA杂交。

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