以Differential display-PCR法選殖綠竹中不同生長時期具差異性表現之基因

Abstract

利用基因差異表現分析法 (DD-PCR) 比較生長中綠竹筍之各組織間基因表現情形。本實驗共獲得 46 個具差異性表現之 DNA 片段，經反向北方分析 (reverse northern analysis) 結果顯示，僅 12 個 DNA 片段具有表現之差異性。其中 AA23100b-2 基因片段經定序與 BLAST 比對分析，預測為 homeoboxes 基因。為選殖出 AA23100b-2 基因全長，進一步以 AA23100b-2 基因片段作為探針，篩選綠竹筍 cDNA 庫。選殖得到之 cDNA 與水稻 putative homeodomain leucine zipper protein 高度同源，並命名為 BoHDZip，其 DNA 序列總長度為1869 bp，但仍缺少基因之 5’端序列。推衍之胺基酸序列含有 homeobox 蛋白質之 START domain 保守性胺基酸。因此推測 BoHDZip 屬於植物 homeobox 蛋白質的 HD-GL2 家族。

Differential display-PCR (DD-PCR) was employed to compare the gene expression profiles among diverse tissues of growing bamboo shoot Bambusa oldhamii. Forty-six differentially expressed fragments were isolated, reamplified, and cloned. Reverse northern analysis showed that only twelve clones exhibited differential expression in patterns in bamboo shoot. The identity of the clone AA23100b-2 was predicted to be homeoboxes gene as revealed by sequencing analysis and BLAST analysis. To clone the full-length cDNA for AA23100b-2, a cDNA library from bamboo shoots was screened by using AA23100b-2 cDNA as probe. A cDNA whose sequence was highly homologous to the homeodomain leucine zipper protein of rice was obtained. This cDNA, designated BoHDZip was 1869bp in length, but still lacted the 5’-end region. The deduced amino acid sequence contained the conserved START domain of homeobox proteins. It might belong to the HD-GL2 family of homeobox protein in plant.