甘蓝型油菜BnCYP78A8的克隆及序列分析  

Cloning and Sequence Analysis of BnCYP78A8 from Brassica napus

作　　者：宋丽丽[1] 贺热情[1] 黄星群[1] 谢敏敏[1] 赵小英[1] 刘选明[1] 

机构地区：[1]湖南大学生物学院,生物能源与材料研究中心,长沙410082

出　　处：《西北植物学报》2012年第4期645-650,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家高技术研究发展计划(2007AA10Z127);国家自然科学基金(30800080)

摘　　要：采用Genome walking方法,首次克隆到甘蓝型油菜BnCYP78A8的基因组序列,根据基因特异性引物克隆到其编码序列。基因组序列长1 679bp,有1个内含子和2个外显子。编码序列长1 605bp,编码534个氨基酸。序列比对分析表明,其氨基酸序列与拟南芥细胞色素P450单加氧酶基因(AtCYP78A8)的相似度高达88%。生物信息学分析显示,该蛋白含有1个细胞色素P450特有的亚铁血红素配合基结合位点和1个跨膜结构域。实时荧光定量PCR分析结果表明,BnCYP78A8在甘蓝型油菜的各个器官组织均有表达,根中表达量最高,表明该基因可能参与根的生长发育。A total of 1 679 bp genomic DNA ot BntCYP78A8 was first cloned trom Brassica napus using Genomic walking method and then the coding sequence was cloned using special primers. The genomic DNA sequence is 1 679 bp,containing one intron and two exons. The coding sequence is l 605 hp,coding for 534 amino acids. Sequence alignment of amino acid showed that BnCYP78A8 shared high identity （88%） with AtCYP78A8 in Arabidopsis thaliana. Bioinformatics analysis indicated that BnCYP78A8 protein contain one cytochrome P450 cysteine berne-iron ligand signature and one transmembrane domain. The real-time PCR results demonstrated that BnCYP78A8 expressed in all indicated tissues of Brassica napus ,and its expression in root was higher than that in other tissues, suggesting that BnCYP78A8 might be involved in root growth.

关 键 词：甘蓝型油菜 BnCYP78A8 GENOME WALKING 序列分析 

分 类 号：Q786[生物学—分子生物学]