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以龙眼松散形胚性愈伤组织为材料,采用RT-PCR结合RACE法克隆Remorin家族5个成员的c DNA全长,Dl Remorin1全长2 127 bp,编码572个氨基酸;Dl Remorin2全长1 829 bp,编码444个氨基酸;Dl Remorin3全长1 937 bp,编码541个氨基酸;Dl Remorin4全长1 743 bp,编码466个氨基酸;Dl Remorin5全长1 043 bp,编码181个氨基酸。Dl Remorin1~5 DNA序列长度分别为:4 054、3 097、3 505、4 690和1 935 bp,所有内含子剪切位点均符合真核生物\\\"GT-AG\\\"规则。所有成员均是不稳定亲水性蛋白,都不具有信号肽,只有Dl Remorin1具有跨膜结构,都具有Remorin家族保守结构域,与其它植物的Remorin具有较高的同源性。系统进化树分析结果表明,Dl Remorin5为经典的Remorin蛋白,Dl Remorin1与Dl Remorin2属于长链Remorin。qRT-PCR结果显示,随着龙眼体胚的发育,Dl Remorin1的表达量呈类\\\"N\\\"状趋势,在心形胚时期和子叶形胚时期较高;Dl Remorin2在心形胚和子叶形胚时期迅速下降;Dl Remorin3在发育中后期迅速升高,并在子叶形胚时期最高;Dl Remorin4在球形胚和鱼雷形胚时期最低,而Dl Remorin5在不完全胚性紧实结构时期最低。说明Dl Remorin1~5在龙眼体胚发生过程的转录水平具有一定的组织特异性和时序性。ps RNA Target预测显示Dl Remorin还可能受到mi RNA家族的调控。
The full-length c DNA of 5 members of Remorin family was cloned by RT-PCR and RACE. The full-length cDNA of Dl Remorin1 was 2 127 bp, encoding 572 amino acids. The full length of Dl Remorin2 was 1829 bp, encoding 444 amino acids; Dl Remorin3 full length 1 937 bp, encoding 541 amino acids; Dl Remorin4 full length 1 743 bp, encoding 466 amino acids; Dl Remorin5 full length 1 043 bp, encoding 181 amino acids. The DNA sequence lengths of Dl Remorin 1-5 were 4 054,3 097,3 505,4 690 and 1 935 bp, respectively. All intron cleavage sites were in accordance with the eukaryote GT-AG rule. All members are unstable hydrophilic proteins and do not have signal peptide. Only Dl Remorin1 has a transmembrane structure and has the Remorin family conserved domain, which has high homology with Remorin of other plants. The results of phylogenetic tree analysis showed that Dl Remorin5 is a classic Remorin protein, and Dl Remorin1 and Dl Remorin2 belong to long-chain Remorin. The results of qRT-PCR showed that with the development of somatic embryos, the expression of Dl Remorin1 was in the \"N\" shape, which was higher in the heart-shaped embryos and in the cotyledons. Dl Remorin2 was found in the heart-shaped and cotyledons Dl Remorin3 rapidly increased in mid and late stages and reached the highest in cotyledon embryos; Dl Remorin4 was the lowest in globular and torpedo embryos, while Dl Remorin5 was the lowest in incomplete embryogenic compacts. Indicating Dl Remorin1 ~ 5 in the longan somatic embryogenesis process has a certain degree of tissue-specific transcriptional level and timing. The prediction of ps RNA Target shows that Dl Remorin may also be regulated by the mi RNA family.