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首页> 外文期刊>Infection and immunity >Insertional inactivation of an intrageneric coaggregation-relevant adhesin locus from Streptococcus gordonii DL1 (Challis).
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Insertional inactivation of an intrageneric coaggregation-relevant adhesin locus from Streptococcus gordonii DL1 (Challis).

机译：戈登链球菌DL1（Challis）中与内聚集相关的粘附素基因座的插入失活。

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Transposon Tn916 was used to insertionally inactivate a coaggregation-relevant locus of Streptococcus gordonii DL1 (Challis). One mutant (F11) was isolated that lost the ability to coaggregate with the streptococcal partners of DL1 but retained the ability to coaggregate with partners belonging to other genera. A probe specific for the region flanking the Tn916 insertion was used to isolate a locus-specific fragment from a chromosomal lambda library. Southern analysis of the resulting phagemids revealed that a 0.5-kb EcoRI fragment hybridized with the F11 probe. Cloning of the 0.5-kb EcoRI fragment into the E. coli-streptococcal insertion vector p(omega) yielded pCW4, which was used to insertionally inactivate the putative coaggregation-relevant gene in DL1. Insertion mutants showed altered coaggregation with streptococci but retained wild-type coaggregation properties with other genera of bacteria. Comparison of immunoblots of cell surface proteins showed a 100-kDa protein in DL1 which was not detected in the Tn916 and pCW4 insertion mutants. These results indicate that the 0.5-kb EcoRI fragment is part of an adhesin-relevant locus that is involved in the production of a 100-kDa protein at the cell surface.
机译：转座子Tn916用于插入灭活戈登链球菌DL1（Challis）的与聚集相关的基因座。分离了一个突变体（F11），该突变体丧失了与DL1链球菌伴侣共聚的能力，但保留了与属于其他属的伴侣共聚的能力。使用对Tn916插入侧翼区域具有特异性的探针从染色体λ文库中分离基因座特异性片段。所得噬菌粒的Southern分析表明0.5kb的EcoRI片段与F11探针杂交。将0.5-kb EcoRI片段克隆到大肠杆菌-链球菌插入载体p（omega）中可得到pCW4，该质粒可用于在灭活DL1中与推定的共聚合相关的基因上失活。插入突变体显示与链球菌的共聚改变，但保留了与其他细菌属的野生型共聚特性。细胞表面蛋白免疫印迹的比较显示DL1中有一个100 kDa的蛋白，在Tn916和pCW4插入突变体中未检测到。这些结果表明，0.5kb的EcoRI片段是与粘附素有关的基因座的一部分，与细胞表面的100kDa蛋白质的产生有关。

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