温州蜜柑线粒体基因的克隆与表达分析

Cloning and Expression Analysis of Mitochondrial Genes from Satsuma Mandarin(Citrus unshiu Marc.)

根据其它植物atp6、cob和coxⅡ基因保守区设计特异引物,利用RT-PCR技术从‘国庆1号’温州蜜柑cDNA中分别扩增出特异性片段,将其克隆至pBluescript(sk+)载体上进行测序。同源性分析表明,所克隆的atp6、cob和coxⅡ与其他植物的对应氨基酸区域同源性分别达到85%、98%和96%以上。RT-PCR分析表明它们在叶片、花瓣以及不同时期的花蕾中都有表达,在幼果中没有表达。Southern分析表明它们在温州蜜柑基因组DNA中为多拷贝。

Synthetic oligonucleotides based on the conserved sequence of atp6, cob and cox Ⅱ of other plants were used to prime the synthesis and amplification of the specific fragments by RT - PCR from Citrus unshiu. PCR products were cloned into pBluescript （sk ＋ ） vector and sequenced. Sequencing analysis showed that the deduced amino acid of atp6, cob and cox Ⅱ shared over 85% , 98% and 96% identities with homologic genes of other plants species respectively. RT - PCR analysis indicated they expressed in the leaf, petal and flower buds during different stages, while not expression in the immature fruit. Southern blot analysis showed that there existed more than one copys in the genome.