言···························································································································1
1.1 研究的目的與意義·······························································································1
1.2宿主與胃腸道微生物的關(guān)系················································································1
1.3 16SrRNA基因檢測(cè)技術(shù)的產(chǎn)生···········································································2
1.4 16SrRNA基因檢測(cè)技術(shù)的應(yīng)用···········································································2
2 材料與方法··············································································································3
2.1 材料·······················································································································3
2.1.1 實(shí)驗(yàn)動(dòng)物············································································································3
2.1.2 生化及分子生物學(xué)試劑····················································································3
2.1.3 腸道基因組DNA提取所需溶液·····································································3
2.1.4 實(shí)驗(yàn)主要器材····································································································4
2.2 方法·······················································································································4
2.2.1羊胚胎胃腸道細(xì)菌分離純化·············································································4
2.2.2 分離細(xì)菌的菌落及形態(tài)特征············································································4
2.2.3 16SrRNA基因同源性分析鑒定分離細(xì)菌·························································5
2.2.3.1 分離細(xì)菌基因組DNA的提取······································································5
2.2.3.2 分離細(xì)菌16SrRNA基因的PCR擴(kuò)增·························································5
2.2.3.3 PCR產(chǎn)物的回收與純化·················································································5
2.2.3.4 16SrDNA文庫(kù)的構(gòu)建與篩選········································································6
2.2.3.5 重組陽(yáng)性克隆的鑒定及測(cè)序········································································7
2.2.3.6 重組陽(yáng)性克隆的酶切及RCR鑒定······························································8
2.2.3.7 16SrRNA基因序列測(cè)定及同源性分析························································8
3 結(jié)果與討論·············································································································9
3.1 結(jié)果······················································································································9
3.1.1 分離細(xì)菌DNA提取·············································································..