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当前位置: 首页 > 文章 > 黑芥ANS基因的克隆和在芸薹属B基因组的PCR鉴别 中国油料作物学报 2013,35 (5) 484-490
Position: Home > Articles > Cloning of ANS gene in Brassica nigra and PCR identification from Brassica B-genomes Chinese Journal of Oil Crop Sciences 2013,35 (5) 484-490

黑芥ANS基因的克隆和在芸薹属B基因组的PCR鉴别

作  者：
严明理;刘丽莉;向建华;丁素萍;舒佳宾;孙婵
单  位：
湖南科技大学生命科学学院
关键词：
黑芥;ANS;克隆;B基因组;PCR鉴别
摘  要：
利用同源克隆方法克隆黑芥ANS基因,获得2个ANS基因拷贝BnANS1和BnANS2,长度分别为1 366bp和1 367bp。这2个基因都有1个76bp的内含子。比较BnANS1和BnANS2基因序列,发现在编码区、5’与3’非编码区和内含子区域都有多态性,5’非编码区有3个碱基的多态性和4个碱基的缺失,编码区有26个碱基的多态性;内含子有2个碱基的多态性;3’非编码区有18个碱基的多态性和3个碱基的插入。这2个ANS拷贝都编码356个氨基酸的蛋白,具有其他物种同样的ANS蛋白保守结构域,属于2OG-Fe(II)加氧酶超家族。BnANS1编码的蛋白质理论分子量为40 448.58Da,等电点为5.05;BnANS2编码的蛋白质理论分子量为40 468.52Da,等电点为5.04。比较这2个ANS基因拷贝编码的蛋白质序列,发现有9个多态性位点。这2个ANS拷贝在黑芥的种皮和胚中均检测到表达。获得了1对ANS引物,能从白菜、黑芥、甘蓝、芥菜型油菜、甘蓝型油菜和埃塞俄比亚芥中,用等位特异PCR方法特异识别来自芸薹属B基因组的ANS基因。
译  名：
Cloning of ANS gene in Brassica nigra and PCR identification from Brassica B-genomes
作  者：
YAN Ming-li;LIU Li-li;XIANG Jian-hua;DING Su-ping;SHU Jia-bin;SUN Chan;School of Life Science,Hunan University of Science and Technology;
关键词：
Brassica nigra;;ANS;;Cloning;;Genome B;;PCR identification
摘  要：
ANS genes were cloned from Brassica nigra by homology-based clone strategy.2 copies named BnANS1 and BnANS2 were obtained.BnANS1 sequence was 1 366bp with a 76bp-intron.BnANS2 sequence gene was 1 367bp with 76bp intron.Sequence analysis showed polymorphisms in coding regions,intron,5'-and 3'-noncoding regions of the two genes,including 3 polymorphic bases and a 4-base deletion in 5'-noncoding region,26 polymorphic bases in coding region,2 polymorphic bases in intron,18 polymorphic bases and a 3-base insertion in 3'-noncoding region.The two copies encoded deduced a polypeptide of 356 amino acids,having the same conserved domain of ANS as other species,belonging to 2OG-Fe(II) oxygenase superfamily.BnANS1 protein was predicted to be 40 448.58Da with pI of 5.05,while BnANS2 was 40 468.52Da with pI of 5.04.Nine polymorphisms were found in their protein sequences.RT-PCR result showed that BnANS1 and BnANS2 expressed in seed coats and embryos in B.nigra.Compared with published ANS gene sequences of Brassica species,a pair of primer was obtained based on B genome-specific nucleotide variations loci,which could effectively identify B-genome origin of ANS in Brassica by allele-specific PCR.

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