龙眼胚性愈伤组织肌动蛋白基因(actin)片段的克隆与序列分析

Cloning and Sequence Analysis of Actin Gene from Dimocarcups Longan Embryogenic Callus

采用同源克隆方法从龙眼胚性愈伤组织分离肌动蛋白基因(actin)。通过保守区和3’RACE扩增,分别获得347bp和837bp的特异片段,经过序列拼接得到895bp的龙眼actin基因3’端序列。与多种植物的肌动蛋白基因进行同源性比较,该片段的核苷酸序列同源性多在80%以上,氨基酸序列同源性大于90%,具有高度保守性。可作为龙眼体胚发生过程中基因表达分析的内参。

The method of Homologous cloning was used to obtain actin gene from Dimocarpus longan embryogenic callus. 347 bp and 837 bp specific fragments were obtained respectively by the amplification of the conservative region and 3＇ end. Stitched the two fragments, 895 bp 3＇end sequence of the actin gene was obtained. By homologous comparing with most plants＇ acitn genes, it shared nucleotide identity more than 80% and amino acid identity more than 90%, which suggests that the longan actin gene is highly conservative and can be used as the internal control for gene expression analysis in longan somatic embryogenesis.