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分别以琯溪蜜柚及其突变体红肉蜜柚(成熟期早20～25d)的成熟果肉cDNA作驱赶子和检测子,利用结合分子镜像选择技术的SSH技术,构建了红肉蜜柚和琯溪蜜柚的正向差减cDNA文库。通过PCR检测cDNA克隆外源插入片段,其大小介于100～1000bp。通过点杂交差异筛选文库,得到102个表达增强2倍或以上的克隆并测序,结果表明这102个克隆代表44个Unigenes,通过序列同源性比对分析,发现获得的Unigenes在功能上主要涉及信号传导、蛋白质合成、应激反应、转运等代谢反应。
The mature cDNA of ripe pomelo was used as the driver and detector of Guanxi honey pomelo and its mutant red meat pomelo (early maturity 20 ~ 25d) respectively. Using the SSH technology combined with molecular image selection technique, the red meat pomelo and Zhengxi pomelo forward subtraction cDNA library. The cDNA inserts were detected by PCR and the size ranged from 100 to 1000 bp. 102 differential clones were screened by dot hybridization and sequenced. The results showed that the 102 clones represented 44 Unigenes. By sequence homology analysis, we found that the obtained Unigenes mainly involved in function Signal transduction, protein synthesis, stress response, transport and other metabolic reactions.