用于多基因表达检测的猕猴桃macroarray阵列

Preparation of kiwifruit macroarray for high throughput gene expression analysis

基因阵列是开展高通量功能基因组学研究的有效手段之一。对适用于多年生果树猕猴桃的macroarray阵列进行了探索,构建了含有42个基因的macroarray阵列,与α-32P标记的cDNA探针杂交产生清晰信号,有效分辨了不同基因或基因家族不同成员的表达差异。DNase处理可去除基因组DNA污染,提高结果准确性。

Gene array is a powerful tool for high throughput gene expression analysis in functional genome research. The present work aims to prepare kiwifruit macroarray membranes and to apply it in expression analysis of ripening-related genes. 42 gene fragments were spotted on positively charged nylon membrane, and hybridized with α-^32P labelled cDNA probes, which generated clear dotted signals with low background. The results showed that these ripening related genes or gene family members had different expression profiles. Furthermore, it was found that removal of potential contamination from genomic DNA by pre-treating total RNA with DNase was essential for the macroarray analysis.