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首页> 外文期刊>Infection and immunity >The Number of Direct Repeats in hagA Is Variable among Porphyromonas gingivalis Strains
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The Number of Direct Repeats in hagA Is Variable among Porphyromonas gingivalis Strains

机译：在牙龈卟啉单胞菌菌株中，hagA中直接重复的数目是可变的

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The coding sequence for the surface protein hemagglutinin A (HagA) of Porphyromonas gingivalis 381 has previously been shown to contain four direct 1.35-kb repeats, designatedrep HA. This study was performed to determine if the number of rep HA units in hagAis consistently 4 or if allelic polymorphism exists among strains and/or upon multiple passage of P. gingivalis. To this end, primers which were homologous to the regions directly 5′ and 3′ of the repeat domain in hagA were synthesized. PCR conditions which allowed amplification of the 8.4-kb repeat region between the primers in P. gingivalis 381 were established. Genomic DNA templates from 13 other P. gingivalis strains and 9 fresh clinical isolates from patients were analyzed under the same conditions as used above. Analysis of these PCR products demonstrated that the strains tested had different numbers (two to four) ofrep HA units in the respective hagAgenes. The PCR products of 8.4, 7.0, and 5.7 kb represent four, three, and two repeats, respectively. One strain from each group (381, four repeats; W83, three repeats; and AJW4, two repeats) was also tested to determine if the number of repeats remained invariant upon passaging onto solid medium. No variability in the number of repeats inhagA within a strain was detected after 18 passages.P. gingivalis 381 was chosen for further testing in a mouse abscess model to determine if conditions of in vivo growth would select for deletions or duplications of the repeated sequences. Five days after infection, no change in the number of repeats was detected in cells recovered from either nonimmunized or preimmunized mice. This data indicates an interstrain variability of the number of repeat units and hence a size variability of the HagA protein of P. gingivalis, but unlike some surface antigens of other pathogenic species, the number of repeats remains relatively stable given the conditions of growth tested here.
机译：先前已显示 Porphyromonas gingivalis 381的表面蛋白血凝素A（HagA）的编码序列包含四个直接的1.35-kb重复序列，称为 rep HA < / sub>。进行这项研究的目的是确定 hagA 中 rep HA 单位的数目是否始终为4，或者菌株之间和/或之后是否存在等位基因多态性。 P的多次通过。牙龈炎。为此，合成了与 hagA 中重复结构域的直接5'和3'区域同源的引物。允许扩增 P中引物之间8.4-kb重复区域的PCR条件。建立了齿龈 381。来自其他13个 P的基因组DNA模板。在与上述相同的条件下，对来自患者的牙龈菌菌株和9株新鲜的临床分离株进行了分析。对这些PCR产物的分析表明，所测试的菌株在各自的 hagA 基因中具有不同数量（2-4个）的 rep HA 单位。 8.4、7.0和5.7 kb的PCR产物分别代表四个，三个和两个重复。还测试了每组的一个菌株（381，四个重复; W83，三个重复;和AJW4，两个重复），以确定在传代到固体培养基上时重复的数目是否保持不变。在18个传代后，在菌株中 hagA 中的重复次数没有变化。选择了龈齿龈炎381在小鼠脓肿模型中进行进一步测试，以确定体内生长条件是否会选择重复序列的缺失或重复。感染五天后，从未免疫或预免疫小鼠中回收的细胞中未检测到重复数的变化。该数据表明重复单元数目的菌株间变异性，并因此指示 P的HagA蛋白的大小变异性。牙龈炎，但与其他病原体的某些表面抗原不同，考虑到此处测试的生长条件，重复次数保持相对稳定。

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