殊异韦荣菌中耐酸相关基因ffh的检测
发布时间：2019-04-15 20:03
【摘要】： 韦荣球菌是口腔中一种革兰阴性专性厌氧球菌，变链球菌胞壁上存在着与韦荣菌结合的受体。韦荣菌对牙面的粘附力较弱，但当牙面上—旦有革兰阳性菌粘附时，韦荣菌随之移入，使菌斑量明显增加，这说明韦荣菌可借助于牙面上的革兰阳性菌而粘附于牙面上，在菌斑形成中也起—定作用。由于韦荣球菌缺乏葡萄糖激酶(glucokinaseu)和果糖激酶(fructokinase)，因此不能代谢碳水化合物和多元醇，但能利用一些碳水化合物代谢的中间产物，特别是利用乳酸盐作为能源。韦荣球菌能将细菌产生的酸性最强的乳酸转变为酸性较弱的乙酸和丙酸，使菌斑pH值上升，所以在牙菌斑生态系和龋病的发生中起重要作用。韦荣菌在酸性环境中能够生长，说明其具有一定的耐酸性。目前在一些原核生物中已分离、测序并克隆了多个耐酸性相关基因。本实验首先在体外培养殊异韦荣菌，用V-gene细菌基因组DNA小量制备试剂盒提取细菌基因组，根据耐酸相关基因ffh的同源性，从最保守区设计一对引物进行PCR，利用pGEM-T载体进行T-A克隆，构建重组质粒，从而检测殊异韦荣菌中是否存在耐酸相关基因ffh，并进行测序鉴定。本实验以殊异韦荣菌为对象，证实殊异韦荣菌中存在耐酸相关基因ffh,对其测序结果进行了同源性比较，结果显示殊异韦荣菌ffh基因序列与GeneBank中登陆的Streptococcus mutan Ffh (ffh)（AE014133.1）UA159相似性100％，，与Streptococcus mutans Ffh (ffh)（AY857409.1）基因相似性99％，与Streptococcus mutans signal recognition particle protein(ffh)（U48883.1，SMU48883）相似性100％，并且所测殊异韦荣菌耐酸相关基因ffh序列已提交美国GenBank，登陆序列号为EU551144。
[Abstract]:Veronica is a gram-negative specific anaerobes in the oral cavity. There is a receptor on the cell wall of Streptococcus mutans that binds to Veronella. The adhesion of Veronica to tooth surface was weak, but when gram-positive bacteria adhered to tooth surface, Weirong bacteria moved into the tooth surface and increased the number of plaque, which indicated that Veronica could adhere to tooth surface with the help of gram-positive bacteria on tooth surface, and when there were Gram-positive bacteria on tooth surface, the bacteria could adhere to tooth surface with the help of Gram-positive bacteria on tooth surface. It also plays a role in plaque formation. Due to the lack of glucose kinase (glucokinaseu) and fructose kinase (fructokinase), Veronica could not metabolize carbohydrates and polyols, but it could use some intermediate products of carbohydrate metabolism, especially lactates as energy sources. Verunococcus can change the acid lactic acid produced by bacteria to acetic acid and propionic acid, which can increase the pH value of plaque, so it plays an important role in the occurrence of dental plaque ecosystem and dental caries. Weirong bacteria can grow in acidic environment, indicating that it has a certain degree of acid tolerance. At present, several acid tolerance related genes have been isolated from prokaryotes, sequenced and cloned. In this experiment, the bacterial genome was extracted by a small amount of genomic DNA of V-gene bacteria in vitro, and a pair of primers were designed from the most conservative region to carry out PCR, according to the homology of the acid tolerance-related gene ffh. The recombinant plasmid was constructed by using pGEM-T vector to clone and construct the recombinant plasmid to detect the existence of acid tolerance-related gene ffh, in Veronica species and to identify the sequence of the gene. In this experiment, the acid-tolerance related gene ffh, was confirmed to be homologous to the result of sequencing, and the results of DNA sequencing were compared with each other in this study, and the results of DNA sequencing were compared with each other. The results showed that the sequence of ffh gene was 100% similar to Streptococcus mutan Ffh (ffh) (AE014133.1 UA159 in GeneBank, 99% to Streptococcus mutans Ffh (ffh) (AY857409.1 gene, and 100% to Streptococcus mutans signal recognition particle protein (ffh) (U48883.1, SMU48883) was 100% similar, and the ffh sequence of acid tolerance-related gene of Veronica vulgaris was submitted to the U.S. GenBank, login serial number EU551144..
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R378
本文编号：2458449
[Abstract]:Veronica is a gram-negative specific anaerobes in the oral cavity. There is a receptor on the cell wall of Streptococcus mutans that binds to Veronella. The adhesion of Veronica to tooth surface was weak, but when gram-positive bacteria adhered to tooth surface, Weirong bacteria moved into the tooth surface and increased the number of plaque, which indicated that Veronica could adhere to tooth surface with the help of gram-positive bacteria on tooth surface, and when there were Gram-positive bacteria on tooth surface, the bacteria could adhere to tooth surface with the help of Gram-positive bacteria on tooth surface. It also plays a role in plaque formation. Due to the lack of glucose kinase (glucokinaseu) and fructose kinase (fructokinase), Veronica could not metabolize carbohydrates and polyols, but it could use some intermediate products of carbohydrate metabolism, especially lactates as energy sources. Verunococcus can change the acid lactic acid produced by bacteria to acetic acid and propionic acid, which can increase the pH value of plaque, so it plays an important role in the occurrence of dental plaque ecosystem and dental caries. Weirong bacteria can grow in acidic environment, indicating that it has a certain degree of acid tolerance. At present, several acid tolerance related genes have been isolated from prokaryotes, sequenced and cloned. In this experiment, the bacterial genome was extracted by a small amount of genomic DNA of V-gene bacteria in vitro, and a pair of primers were designed from the most conservative region to carry out PCR, according to the homology of the acid tolerance-related gene ffh. The recombinant plasmid was constructed by using pGEM-T vector to clone and construct the recombinant plasmid to detect the existence of acid tolerance-related gene ffh, in Veronica species and to identify the sequence of the gene. In this experiment, the acid-tolerance related gene ffh, was confirmed to be homologous to the result of sequencing, and the results of DNA sequencing were compared with each other in this study, and the results of DNA sequencing were compared with each other. The results showed that the sequence of ffh gene was 100% similar to Streptococcus mutan Ffh (ffh) (AE014133.1 UA159 in GeneBank, 99% to Streptococcus mutans Ffh (ffh) (AY857409.1 gene, and 100% to Streptococcus mutans signal recognition particle protein (ffh) (U48883.1, SMU48883) was 100% similar, and the ffh sequence of acid tolerance-related gene of Veronica vulgaris was submitted to the U.S. GenBank, login serial number EU551144..
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R378
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