荔枝胚败育差异表达基因cDNA 片段的克隆及序列分析

 利用抑制消减杂交(Suppressive subtraction hybridization , SSH) 克隆荔枝败育胚的差异表达基因cDNA 片段。分别以荔枝‘桂味’正常发育胚为driver (驱赶子) , 败育胚为tester (检测子) , 建立差异表达cDNA 文库, 代表桂味败育胚特异表达的cDNA。经Virtual Northern 检测, 在桂味正常发育胚/ 败育胚差异表达cDNA 文库中得到3 个阳性克隆, 序列分析表明这3 个克隆对应的两个序列在荔枝中为首次报道。

Cloning and Sequencing of cDNA Fragments Differentially Expressed between Aborted and Normal Development Embryo in Litchi

The suppressive subtraction hybridization (SSH) technique was used to clone the cDNA fragments differentially expressed between aborted and normal development embryo of ‘Guiwei’varieties. One positive differential expressing cDNA library were constructed using cDNAs of normal development embryo of Guiwei as driver and the cDNAs of aborted embryo of Guiwei varieties as tester. This cDNA library indicated the differential expressingcDNA fragments in Guiwei aborted embryo and Guiwei normal development embryo. By Virtual Northern blots, 3 positive clones were obtained. Subsequently sequencing analysis showed the 3 clones were corresponding 3 knew gene sequence and the 3 cDNA fragments were first reported in litchi varieties.