大白菜碳酸酐酶基因的克隆与序列分析  

Cloning and Sequence Analysis of Carbonic Anhydrase Gene in Chinese Cabbage

作　　者：赵会芳[1] 巩振辉[1] 赵利民[1] 柯桂兰[1] 

机构地区：[1]西北农林科技大学园艺学院,陕西杨陵712100

出　　处：《西北植物学报》2010年第9期1728-1732,共5页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(30771467);陕西省"13115"科技创新工程重大科技专项(2008ZDKG-03);陕西省蔬菜产业技术体系

摘　　要：以大白菜雄性不育系和保持系花蕾差异表达片段EST H9为信息探针,在GenBank数据库中进行同源EST序列检索,并对亲缘关系近的同源EST序列进行拼接,得到大白菜EST H9的5-′cDNA序列。根据拼接组装所得的5-′cDNA序列,进行3-′RACE引物的设计,经RACE扩增、测序,获得了大白菜α-碳酸酐酶3基因的cDNA全长序列并将其登录到GenBank(登录号为GU143061),命名为BrACA3。该cDNA全长998 bp,编码270个氨基酸。同源分析显示该cDNA序列推导的氨基酸序列与拟南芥α-碳酸酐酶3一致性达78%。氨基酸序列分析表明,该蛋白具备跨膜功能,在第19和20个氨基酸之间存在一个信号肽序列,存在丝氨酸、苏氨酸和酪氨酸磷酸化位点。在大白菜花蕾败育过程中α-碳酸酐酶3基因不表达,只在保持系B7的大花蕾时期表达。A differential TDF（Transcript derived fragments） EST H9 from the RNA of male sterile line RC7 and its maintainer line of Chinese cabbage was used as a querying probe to blast homologous ESTs sequences in the GenBank database,we spliced the homologous EST sequences assembled from GenBank,and gained a 5′-cDNA sequence.Then,the primer of 3′-RACE was designed,the full length cDNA of α-carbonate dehydratase 3 gene in Chinese cabbage was gained through RACE amplifying,and sequenced and registered as accession number of GU143061 in GenBank,named BrACA3,which is 998 bp length and coded 270 amino acids.Homologous analysis showed that amino acid sequence derived from it had 78% identity with α-carbonate dehydratase 3（ACA3） in Arabidopsis.Amino acid sequence analysis showed that the ACA3 is a transmembrane protein and there is a signal peptide sequence between 19 and 20 amino acid,there are serine,threonine and tyrosine phosphorylation sites.ACA3 gene in Chinese cabbage was not expressed during bud aborting,and was expressed in large flower buds of B7.

关 键 词：大白菜 细胞质雄性不育 α-碳酸酐酶3 

分 类 号：Q785[生物学—分子生物学]