小立碗藓细胞色素P450基因PpCYP51G1的分离及转化载体构建

Isolation of Cytochrome P450 Gene PpCYP51G1 from Physcomitrella patens and Construction of Its Transformation Vectors

从小立碗藓(Physcomitrella patens)中克隆了细胞色素P450 CYP51基因PpCYP51G1的全长编码序列,编码区长1 509 bp,预测编码488个氨基酸,蛋白二级结构含有自由卷曲、伸展片段和α-螺旋,N端含有一个跨膜区域。构建了PpCYP51G1的功能缺失载体,并利用PEG介导的原生质体转化技术获得该基因的功能缺失转化体;同时,构建了PpCYP51G1-GFP融合蛋白载体,亚细胞定位结果表明,PpCYP51G1定位在内质网中。

The full length cDNA sequence of cytochrome P450CYP51 gene named as PpCYP51G1 was cloned from Physcomitrella patens. Its code region was 1 509 bp, and it encoded 488 amino acids. The PpCYP51 G1 protein contained one transmembrane region at the N end, and its secondary structure contained random coils, extended strands and alpha helixes. The knockout vector for PpCYP51G1 was constructed and the loss - of - function transformants were obtained. The expression vector for PpCYP51G1 - GFP fusion was also constructed to investigate the subcellular localization of PpCYP51G1, and the result showed that PpCYP51G1 expressed in endoplasmic reticulum.