胡萝卜DcPAB基因的分离及其结构与功能分析  

Isolation and Functional Identification of DcPAB from Daucus carota

作　　者：林慧馨[1] 张雷[1] 杨志攀[1] 黄美娟[2] 吴乃虎[1] 

机构地区：[1]中国科学院遗传与发育研究所,北京100080 [2]北京大学生命科学学院,北京100871

出　　处：《中国生物化学与分子生物学报》2004年第3期319-324,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金资助课题 (No .3 963 0 0 40 )~~

摘　　要：运用改进的减法杂交技术分离到胡萝卜Poly(A)结合蛋白基因DcPAB .其cDNA编码区长度为 1 977bp ,编码 6 5 8个氨基酸和 1个终止密码子 .基因组转录序列区长度为 4 6 1 6bp ,包含 9个外显子和 8个内含子 .DcPAB在胡萝卜基因组中为单拷贝基因 .该基因在胡萝卜体细胞胚中特异性表达 ,且其表达活性在调控 解调控前后有明显差异 .体外结合实验表明 ,在大肠杆菌中表达并纯化的DcPAB蛋白具有与oligo(A) 2 0 特异性结合的性能 .酵母突变体互补实验进一步证明 。Using modified virtual subtraction method, a poly(A) binding protein gene, DcPAB , had been isolated from carrot somatic embryo. The coding region of DcPAB contained 1977 base pairs, coding 658 amino acids and a stop codon. The full length transcript was 4616 bp in length, consisting of 9 exons and 8 introns. DcPAB was a single gene in carrot genome. As shown by Northern blotting, DcPAB was detected only in carrot somatic embryo, and its expression displaied distinct difference between the regulated and deregulated embryos. With gel retardation experiment, it was confirmed that the recombinant DcPAB was able to bind oligo(A) 20 specifically. In vivo analysis revealed that DcPAB could complement the deletion of the PAB in yeast pab1 mutant.

关 键 词：胡萝卜体细胞胚 DcPAB 凝胶阻滞检测 

分 类 号：Q291[生物学—细胞生物学] Q786