【摘要】： 韋榮球菌是口腔中一種革蘭陰性專(zhuān)性厭氧球菌，變鏈球菌胞壁上存在著與韋榮菌結(jié)合的受體。韋榮菌對(duì)牙面的粘附力較弱，但當(dāng)牙面上—旦有革蘭陽(yáng)性菌粘附時(shí)，韋榮菌隨之移入，使菌斑量明顯增加，這說(shuō)明韋榮菌可借助于牙面上的革蘭陽(yáng)性菌而粘附于牙面上，在菌斑形成中也起—定作用。由于韋榮球菌缺乏葡萄糖激酶(glucokinaseu)和果糖激酶(fructokinase)，因此不能代謝碳水化合物和多元醇，但能利用一些碳水化合物代謝的中間產(chǎn)物，特別是利用乳酸鹽作為能源。韋榮球菌能將細(xì)菌產(chǎn)生的酸性最強(qiáng)的乳酸轉(zhuǎn)變?yōu)樗嵝暂^弱的乙酸和丙酸，使菌斑pH值上升，所以在牙菌斑生態(tài)系和齲病的發(fā)生中起重要作用。韋榮菌在酸性環(huán)境中能夠生長(zhǎng)，說(shuō)明其具有一定的耐酸性。目前在一些原核生物中已分離、測(cè)序并克隆了多個(gè)耐酸性相關(guān)基因。本實(shí)驗(yàn)首先在體外培養(yǎng)殊異韋榮菌，用V-gene細(xì)菌基因組DNA小量制備試劑盒提取細(xì)菌基因組，根據(jù)耐酸相關(guān)基因ffh的同源性，從最保守區(qū)設(shè)計(jì)一對(duì)引物進(jìn)行PCR，利用pGEM-T載體進(jìn)行T-A克隆，構(gòu)建重組質(zhì)粒，從而檢測(cè)殊異韋榮菌中是否存在耐酸相關(guān)基因ffh，并進(jìn)行測(cè)序鑒定。本實(shí)驗(yàn)以殊異韋榮菌為對(duì)象，證實(shí)殊異韋榮菌中存在耐酸相關(guān)基因ffh,對(duì)其測(cè)序結(jié)果進(jìn)行了同源性比較，結(jié)果顯示殊異韋榮菌ffh基因序列與GeneBank中登陸的Streptococcus mutan Ffh (ffh)（AE014133.1）UA159相似性100％，，與Streptococcus mutans Ffh (ffh)（AY857409.1）基因相似性99％，與Streptococcus mutans signal recognition particle protein(ffh)（U48883.1，SMU48883）相似性100％，并且所測(cè)殊異韋榮菌耐酸相關(guān)基因ffh序列已提交美國(guó)GenBank，登陸序列號(hào)為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..
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類(lèi)號(hào)】：R378

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