光叶楮肌动蛋白基因片段的克隆及其序列分析

Cloning and Sequence Analysis of Actin Gene from Broussonetia papyrifera

目的:克隆构树Actin基因,为在分子生物学中研究外源基因在构树中的表达提供内参,为克隆和分析桑科其它植物的actin基因提供参考。方法:根据GenBank中一桑科植物桑树Actin基因的EST序列,设计引物,提取构树叶片总RNA,通过RT-PCR克隆目标片段。结果:获得一段大小为238bp的基因片段,经测序、同源性比较,这条片段的EST序列与桑树Actin基因、巴旦杏Actin基因的核苷酸同源性分别为96%、92%;氨基酸序列的同源性则分别为100%、98.73%。结论:通过实验结果分析表明获得了构树actin基因EST序列。

Objective： Clone Broussonetia papyrifera＇s actin gene, so thai it can be used as internal standard when study foreign genes＇ expression in Broussonetia papyrifera and provide available information for cloning and analyzing other Moraceae plants＇ actin gene. Methods： Primer was designed according to one kind of Moraceae plants＇ s （ Morus alba ） acfin EST. The tolal RNA was isolated from the leaf of Broussonetia papyrifera, the RT- PCR was used to clone the gene. Results： 238bp nucleotides sequence was obtained, as homologous comparing with the EST anti amino acid sequences of Morus albal and Prunus dulcis, it shared nucleotide identity of 96% and 92% respectively and ,amino acid identity of the 100% and 98.7%, respectively. Conclusion： The obtained nucleotides sequence is Broussonetia papyrifera＇s actin gene.