大豆食心虫28SrDNA基因的克隆及表达分析

Cloning and characterization of 28SrDNA gene from soybean pod borer(Leguminivora glycinivorella Matsumurav)

文章克隆并分析鳞翅目大豆食心虫(Leguminivora glycinivorellaMatsumurav)28S核糖体RNA基因(28SrDNA)全序列。系统进化树分析表明,大豆食心虫28SrDNA与其他鳞翅目昆虫的28SrDNA同源性较高。利用荧光定量PCR对28SrDNA在大豆食心虫不同生育时期和幼虫不同组织的表达量进行分析。结果表明,28SrDNA基因在成虫期表达量最高,卵中表达量最低;在幼虫脂肪体和睾丸的表达量最高,中肠和卵巢的表达量最低。为进一步干扰大豆食心虫28SrDNA基因表达,将28SrDNA序列片段亚克隆到RNAi载体L4440中,经PCR和酶切鉴定证明,成功构建表达大豆食心虫28SrDNA dsRNA的RNAi载体L4440-28SrDNA。

The entire sequence of the 28S ribosomal RNA gene（28SrDNA） of the soybean pod borer（Leguminivora glycinivorella Matsumurav） was cloned and sequenced.Phylogenetic tree analysis showed that the sequences 28SrDNA was highly homology with the 28SrDNA of other epidopteranspecies.qPCR experiments was carried out to analyze that the 28SrDNA transcript was expressed in the throughout feeding stage,with a higher expression level in the adult,while a lower expression level of 28rDNA was observed in the egg.The RT-PCR analyses also showed that 28SrDNA was expressed highly at the transcript level in the ovary and fatbod,but lower expression in the midgut and testis.28SrDNA were obtained.