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黑麦草中木质素的结构、含量是决定其饲用品质的主要因素。漆酶是木质素生物合成中的关键酶之一,属于多铜氧化酶基因家族。根据报道的植物漆酶编码基因的氨基酸序列中铜离子结合域及N-端糖基化位点两个保守区设计4对引物,以黑麦草总DNA为模板,经PCR扩增、克隆、测序后得到3个有效序列,分别命名为Lac10-1、Lac8-1和Lac11-1。序列分析表明,3个基因片段的编码蛋白不仅含有保守的蓝铜氧化酶结合区域HAH和N-末端糖基化位点,而且分别与黄杨、黑麦草中分离的漆酶编码蛋白的氨基酸序列具有较高的同源性。因而,推断克隆到的3个基因片段是黑麦草漆酶基因家族的成员。
The structure and content of lignin in ryegrass are the main factors that determine its forage quality. Laccase is one of the key enzymes in lignin biosynthesis and belongs to the multi-copper oxidase gene family. Four pairs of primers were designed according to the two conserved regions of the copper ion-binding domain and the N-terminal glycosylation site in the amino acid sequence of the reported plant laccase-encoding gene. The total DNA of ryegrass was used as a template to amplify, clone and sequence After 3 valid sequences were obtained, named Lac10-1, Lac8-1 and Lac11-1. Sequence analysis showed that the encoded proteins of the three genes contained not only the HAH and N-terminal glycosylation sites of the conserved blue copper oxidase binding region but also the amino acid sequences of the laccase-encoding proteins isolated from the genus Populus and ryegrass, respectively High homology. Thus, it is concluded that the three cloned genes are members of the ryegrass laccase gene family.