杨树幼茎特异表达基因及PsnLAC基因的克隆

摘要

利用cDNA-AFLP技术对小黑杨茎、叶基因的差异表达进行分析.结果表明,64对引物组合在茎和叶中共扩增出4 192条带,其中差异条带2 275条.通过对茎中特异表达基因的克隆和测序,获得了4条cDNA序列.经Blast-x比对分析表明,它们分别是类伸展蛋白、漆酶、过氧化物酶和细胞壁相关水解酶.进行RT-PCR分析发现,4个基因在茎中的表达水平明显高于叶和根.其中,以漆酶的基因片段为基础进行生物信息学分析和设计引物,用RT-PCR技术克隆到了1 413 bp包含开放阅读编码框的cDNA片段,其开放阅读编码框长度为1 020bP,编码339个氨基酸残基,与来自其他植物漆酶的氨基酸序列具有较高的同源性.%Different expressed genes between stems and leaves of Populus simonii × P. nigra were observed by the cDNA-AFLP method. Result showed that a total of 4 192 fragments were amplified from the cDNA of stems and leaves, and a total of 2275 fragments were differential displayed genes between stems and leaves. Four fragments from stems were cloned and sequenced.Blast-x analysis showed that the four clones were extension like protein, laccase, peroxidase, cell wall-associated hydrolase.Based on a cDNA fragment of laccase, the 1413 bp cDNA of laccase in Populus simonii × P. nigra was obtained using RT-PCR,including a 1 020 bp open reading frame, and encoding 339 amino acid residues sharing higher homology with other plants.