Development and evaluation of whole-genome oligonucleotide array for Acidithiobacillus ferrooxidans ATCC 23270  

作　　者：李乾 申丽 罗海良 尹华群 廖立琴 邱冠周 刘学端 

机构地区：[1]Key Laboratory of Biometallurgy of Ministry of Education,School of Minerals Processing and Bioengineering,Central South University

出　　处：《中国有色金属学会会刊：英文版》2008年第6期1343-1351,共9页Transactions of Nonferrous Metals Society of China

基　　金：Project(50621063)supported by the National Natural Science Foundation of China;Project(2004CB619201)supported by the National Basic Research Program of China

摘　　要：为了有效地在 whole-genomic 监视 Acidithiobacillus ferrooxidans ATCC 23270 的特征，铺平，整个染色体的 50-mer-based oligonucleotide microarray 基于 A 的 3 217 ORF 被开发。ferrooxidans ATCC 23270 染色体。基于人工的 oligonucleotide 探针，结果证明最佳的杂交温度是 45 ??????? 汯杩湯 ' 拙船樐??????To effectively monitor the characteristic of Acidithiobacillus ferrooxidans ATCC 23270 at the whole-genomic level, a whole-genome 50-mer-based oligonucleotide microarray was developed based on the 3 217 ORFs of A. ferrooxidans ATCC 23270 genome. Based on artificial oligonucleotide probes, the results showed that the optimal hybridization temperature was 45 ℃. Specificity tests with the purified PCR amplifications of 5 genes （Sulfide-quinone reductase, Cytochrome C, Iron oxidase, Mercuric resistance protein, Nitrogenase iron protein） of A. ferrooxidans ATCC 23270 indicated that the probes on the arrays appeared to be specific to their corresponding target genes. Based on the WGA hybridization to global transcriptional difference of A.ferrooxidans ATCC 23270 strains cultured with Fe（ Ⅱ ） and S（0）, the developed 50-mer WGA could be used for global transcriptome analysis ofA. ferrooxidans ATCC 23270. The detection limit was estimated to be approximately 5 ng with the genomic DNA, and at 100 ng of the DNA concentration, all of the signals reached the saturation. In addition, strong linear relationships were observed between hybridization signal intensity and the target DNA concentrations （r^2=0.977 and 0.992）. The results indicated that this technology had potential as a specific, sensitive and quantitative tool for detection and identification of the strain A.ferrooxidans ATCC 23270 at the whole-genome level.

关 键 词：寡核苷酸数列 ATCC 23270 特异性 灵敏性 量化 

分 类 号：TF18[冶金工程—冶金物理化学]