| 黑芥ANS基因的克隆和在芸薹属B基因组的PCR鉴别 |
| |
| 引用本文： | 严明理,刘丽莉,向建华,丁素萍,舒佳宾,孙婵.黑芥ANS基因的克隆和在芸薹属B基因组的PCR鉴别[J].中国油料,2013(5):484-490. |
| |
| 作者姓名： | 严明理 刘丽莉 向建华 丁素萍 舒佳宾 孙婵 |
| |
| 作者单位： | 湖南科技大学生命科学学院,湖南湘潭411201 |
| |
| 基金项目： | 国家自然科学基金（31000723）;湖南省高校青年骨干教师项目（湘教通2011-388） |
| |
| 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BnAⅣS1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.
|
| 关 键 词： | 黑芥 ANS 克隆 B基因组 PCR鉴别 |
Cloning of ANS gene in Brassica nigra and PCR identification from Brassica B-genomes |
| |
| Authors: | YAN Ming-liSchool of Life Science Hunan University of Science and Technology Xiangtan China LIU Li-liSchool of Life Science Hunan University of Science and Technology Xiangtan China XIANG Jian-huaSchool of Life Science Hunan University of Science and Technology Xiangtan China DING Su-pingSchool of Life Science Hunan University of Science and Technology Xiangtan China SHU Jia-binSchool of Life Science Hunan University of Science and Technology Xiangtan China SUN Chan |
| |
| Affiliation: | YAN Ming-li(School of Life Science, Hunan University of Science and Technology, Xiangtan 411201, China) LIU Li-li(School of Life Science, Hunan University of Science and Technology, Xiangtan 411201, China) XIANG Jian-hua(School of Life Science, Hunan University of Science and Technology, Xiangtan 411201, China) DING Su-ping(School of Life Science, Hunan University of Science and Technology, Xiangtan 411201, China) SHU Jia-bin(School of Life Science, Hunan University of Science and Technology, Xiangtan 411201, China) SUN Chan(School of Life Science, Hunan University of Science and Technology, Xiangtan 411201, China) |
| |
| Abstract: | 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. |
| |
| Keywords: | Brassica nigra ANS Cloning Genome B PCR identification |
| 本文献已被 维普 等数据库收录！ |
|
