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漆酶是木质素生物合成途径的关键酶,其高度保守的铜离子结合区以及N末端糖基化位点使利用同源克隆技术成为可能。本研究利用GeneBank中公布的黑麦草漆酶基因全长cDNA序列及在玉米数据库中的比对结果设计特异引物,以8份AS系列玉米自交系为模板进行PCR筛选。经过提取质粒、测序及对推导的氨基酸序列进行比对,发现所克隆基因片段中含有保守的H-A-H基元和N末端糖基化位点,为以后玉米漆酶基因全长的克隆以及饲用玉米改良打下了基础。
Laccase is a key enzyme in the lignin biosynthesis pathway, and its highly conserved copper ion binding sites and N-terminal glycosylation sites make it possible to utilize homologous cloning techniques. In this study, specific primers were designed based on the full-length cDNA sequence of ryegrass laccase gene published in GeneBank and the comparison results in maize database. Eight AS inbred lines were used as templates for PCR screening. After the plasmid was extracted, sequenced and the deduced amino acid sequence was compared, it was found that the cloned gene fragment contains the conserved HAH motif and N-terminal glycosylation site, which is the full-length clone of maize laccase gene and maize Improve laid the foundation.