葛根膨大相关基因表达的cDNA-AFLP分析

cDNA-AFLP Analysis on Genes Associated with Tuberous Root Swelling in Pueraria lobata(Wild.) Ohwi

为从本质上揭示葛块根膨大的分子机理,将cDNA-AFLP技术运用于葛块根膨大的野生型和突变型发育过程的基因表达差异的分析,筛选出5个特异表达片段,其中野生型Y0、Y1和突变型T0、T14个成功测序,结果显示:DNA片段大小分别为384、248、241和161 bp;BLAST搜索结果表明:Y0与大豆假拟蛋白相关的未知mRNA同源性99%;Y1与罗伊乳杆菌木糖异构酶基因序列同源性95%;T0与豇豆小分子热休克蛋白mRNA序列同源性87%;T1与番茄的类胡萝卜素异构酶基因同源性98%;Y0翻译的蛋白质与拟南芥亚种赤霉素结合蛋白β-2亚基有一定同源性,Y1和T0、T1翻译的蛋白质在核酸蛋白数据库中均未搜索到同源序列。

In order to essentially reveal the molecular mechanism of Pueraria lobata（Wild.） Ohwi tuberous root swelling,this study adopts cDNA-AFLP to analyse the gene expression differences in the swollen wild type and mutant type of Pueraria lobata（Wild.） Ohwi tuberous root in its developing process,and has selected five special clips,four DNA fragment sequences,including wild types Y1 and Y0,mutation types T0 and T1.The results show： the length of the DNA fragments are 384,248,241 and 161 bp respectively;and BLAST search results show that Y0 is associated with unknown mRNA of hypothetical protein in soybean with identity of 99%;Y1 is associated with Roy lactobacillus xylose isomerase gene sequences with identity of 95%;T0 is associated with cowpea small heat shock protein mRNA sequence with identiy of 87%;T1 is associated with tomato carotenoids isomerase gene homology with identity of 98%;in nucleotide-Protein database,the protein of T1 is associated with gibberellic acid binding proteins subunit β-2 in arabidopsis subspecies with some identity;but the significant homologous protein sequences translated by T0,Y0 and Y1 have not been found in the databases yet.