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

作　　者：丁谦[1,2] 叶帅[1,3] 李景娟[1] 朱正歌[3] 高建伟[1] 

机构地区：[1]山东省农业科学院蔬菜研究所/山东省设施蔬菜生物学重点实验室/国家蔬菜改良中心山东分中心,山东济南250100 [2]甘肃农业大学,甘肃兰州730070 [3]河北师范大学生命科学学院,河北石家庄050016

出　　处：《山东农业科学》2013年第10期1-6,11,共7页Shandong Agricultural Sciences

基　　金：国家自然科学基金(30671177);山东省优秀中青年科学家科研奖励基金(2006BS06001);山东省农业科学院博士基金(2006-2009)资助

摘　　要：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.

关 键 词：小立碗藓 细胞色素P450 CYP51 克隆 转化载体构建 

分 类 号：Q781[生物学—分子生物学]