頁籤選單縮合
題名 | 以非放射性核酸探針定量環境中的脫氮菌(1)--核酸探針的篩選=Quantification of Soil Denitrifiers by Non-Isotope Nucleic Acid Probe Part I. Screen of Nucleic Acid Probe for Denitrifiers |
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作者姓名(中文) | 吳秋曄; 梁志欽; | 書刊名 | 健康與照顧科學學刊 |
卷期 | 1:1 2012.02[民101.02] |
頁次 | 頁131-143 |
分類號 | 414.83 |
關鍵詞 | 脫氮菌; 亞硝酸還原酵素; 核酸探針; Denitrifier; Nitrite reductase; Nucleic acid probe; |
語文 | 中文(Chinese) |
中文摘要 | 本研究之目的在篩選分別能與具有cytochrome c,d1 型或Cu 型亞硝酸還原酵素的脫氮菌 雜交的核酸探針。該核酸探針是以基因序列(5'→ 3')分別為GTC AAG GTG GAA CTG GTC AA 與CCT GTT GGC TTG CGA ATG AA 作為聚合酵素連鎖反應中的引子,14 株具有Cu 型 亞硝酸還原酵素的脫氮菌之DNA 做為模板,進行聚合酵素連鎖反應後,純化聚合酵素連鎖 反應產物做為核酸探針,分別與實驗菌株進行雜交試驗。雜交結果顯示,以Alcaligenes eutrophus BCRC 13036 之DNA 為模板,Mg2+濃度為1.2 mM,鍊合溫度為42.5℃,複製出約 781 bp 的Cu 型亞硝酸還原酵素的基因片段,作為核酸探針,可與具有cytochrome c,d1 型或 Cu 型亞硝酸還原酵素的多種不同屬之脫氮菌雜交。 |
英文摘要 | A DNA probe for determination of denitifiers was screened in the present study. We synthesized a pair of primers for the polymerase chain reaction, the 5'→ 3' sequences of primers separately were GTC AAG GTG GAA CTG GTC AA and CCT GTT GGC TTG CGA ATG AA. The templates of the polymerase chain reaction were used the DNAs from the 14 strains of denitrifiers containing copper-type nitrite reducatae. Under the following amplification conditions, 1.2 mM Mg2+, 42.5℃ annealing temperature for the primers were required. The amplified fragments were purified to used as probes. The probes were labeled by digoxigenin-dUTP. Further confirmation of specificity to denitrifying bacteria for probes was performed by dot blotting hybridization. The results showed that the 781-bp fragment of Alcaligenes eutrophus BCRC 13036 detected more genera of denitrifiers containing copper-type or cytochrome c,d1 nitrite reductase than other probes. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。