發(fā)布時(shí)間：2018-03-04 01:11

  本文選題：布魯氏菌病　切入點(diǎn)：布魯氏菌　出處：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：為了鑒別布魯氏菌S2疫苗株與其它布魯氏菌,本研究建立了鑒別布魯氏菌S2疫苗株的斑點(diǎn)雜交法。針對(duì)布魯氏菌S2疫苗株IclR基因序列保守區(qū)與其他菌株的差異,設(shè)計(jì)合成針對(duì)S2疫苗株的大小為29 bp的寡核苷酸探針,并進(jìn)行地高辛標(biāo)記。根據(jù)探針作用位點(diǎn)設(shè)計(jì)引物,對(duì)擴(kuò)增的PCR產(chǎn)物經(jīng)過回收、純化、定量,變性后經(jīng)過120℃30 min烘烤固定在NC膜上,然后與探針雜交、顯色。結(jié)果顯示,本實(shí)驗(yàn)設(shè)計(jì)的PCR引物1能夠?qū)2疫苗株擴(kuò)增出約330 bp的核酸片段,對(duì)其他參考布魯氏菌擴(kuò)增出約360 bp的核酸片段；采用末端標(biāo)記法可以成功地將地高辛連接到本研究設(shè)計(jì)合成的寡核苷酸探針上,標(biāo)記效果良好；探針只能和S2的PCR產(chǎn)物雜交,特異性強(qiáng),重復(fù)性好,最低能夠檢測到10 pg的PCR片段,能夠在10h內(nèi)鑒別出布魯氏菌S2疫苗株。
[Abstract]:In order to identify Brucella S2 vaccine strain from other brucella strains, a dot hybridization method was established to identify the Brucella S2 vaccine strain. The difference of IclR gene conserved region between Brucella S2 vaccine strain and other strains was studied. A 29bp oligonucleotide probe was designed and synthesized for S2 vaccine strain and labeled with digoxigenin. Primers were designed according to the action site of the probe to recover, purify and quantify the amplified PCR products. After denaturation, the NC membrane was roasted at 120 鈩，

本文編號(hào)：1563484