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通过脉冲场凝胶电泳研究犬胃螺杆菌、比氏螺杆菌和所罗门螺杆菌的遗传多样性。

Genetic diversity of canine gastric helicobacters, Helicobacter bizzozeronii and H. salomonis studied by pulsed-field gel electrophoresis.

作者信息

HäUNNINEN M-L, Hirvi U

出版信息

J Med Microbiol. 1999 Apr;48(4):341-347. doi: 10.1099/00222615-48-4-341.

Abstract

Genetic diversity of Helicobacter bizzozeronii and H. salomonis, two recently identified canine gastric Helicobacter spp., was studied by pulsed-field gel electrophoresis (PFGE). All 15 Finnish H. bizzozeronii strains collected between 1991 and 1996 from pet dogs produced different PFGE patterns with all restriction endonucleases studied (AscI, ApaI, SpeI, NotI and PacI) suggesting significant genetic diversity. The five independent H. salomonis strains produced four different patterns with these enzymes; two strains showed identical patterns with all the enzymes. Three separate isolates from one dog had identical patterns, suggesting long-lasting infection with the same strain. H. salomonis strains had several small fragments common for all strains, suggesting relatedness. The PFGE method was shown to be useful for epidemiological studies of canine gastric helicobacter infection. Hybridisation of the DNA digests with digoxigenin-labelled ureB or 16S rRNA gene probes generated by PCR indicated conservation in the localisation of these genes in the H. salomonis genome, because the probes hybridised with similar size fragments of different strains. In contrast, the probes hybridised with different size fragments of H. bizzozeronii strains. Comparison of Southern blots of PFGE patterns digested with SpeI, ApaI and AscI indicated that each species has two 16S rRNA genes and one urease gene. Genome sizes of 11 H. bizzozeronii strains estimated from SpeI and NotI patterns were c. 1.6-1.9 Mb and those of five H. salomonis strains estimated from NotI and PacI patterns were c. 1.7-1.8 Mb.

摘要

采用脉冲场凝胶电泳(PFGE)技术,对最近发现的两种犬胃幽门螺杆菌——比氏幽门螺杆菌(Helicobacter bizzozeronii)和所罗门幽门螺杆菌(H. salomonis)的遗传多样性进行了研究。1991年至1996年间从宠物狗身上采集的所有15株芬兰比氏幽门螺杆菌菌株,在用所有研究的限制性内切酶(AscI、ApaI、SpeI、NotI和PacI)进行分析时,均产生了不同的PFGE图谱,这表明存在显著的遗传多样性。5株独立的所罗门幽门螺杆菌菌株在用这些酶分析时产生了4种不同的图谱;2株菌株在用所有酶分析时显示出相同的图谱。从一只狗身上分离出的3个单独菌株具有相同的图谱,这表明该菌株存在长期感染。所罗门幽门螺杆菌菌株有几个所有菌株共有的小片段,这表明它们之间存在相关性。PFGE方法被证明可用于犬胃幽门螺杆菌感染的流行病学研究。用PCR产生的地高辛标记的ureB或16S rRNA基因探针与DNA酶切片段杂交,表明这些基因在所罗门幽门螺杆菌基因组中的定位具有保守性,因为探针与不同菌株的相似大小片段杂交。相比之下,探针与比氏幽门螺杆菌菌株的不同大小片段杂交。用SpeI、ApaI和AscI酶切的PFGE图谱的Southern印迹比较表明,每个物种都有两个16S rRNA基因和一个脲酶基因。根据SpeI和NotI图谱估计的11株比氏幽门螺杆菌菌株的基因组大小约为1.6 - 1.9 Mb,根据NotI和PacI图谱估计的5株所罗门幽门螺杆菌菌株的基因组大小约为1.7 - 1.8 Mb。

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