黄瓜异戊烯基转移酶基因的克隆及序列分析  

Cloning and Sequence Analysis of Isopentenyltransferases Gene from Cucumis sativus

作　　者：张微微[1,2] 潘俊松[1] 刘淳中[1] 何欢乐[1] 蔡润[1] 

机构地区：[1]上海交通大学农业与生物学院,上海200240 [2]中国科学院上海辰山植物科学研究中心,上海201602

出　　处：《上海交通大学学报（农业科学版）》2010年第6期487-491,498,共6页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：国家自然科学基金(30671111)

摘　　要：根据GenBank拟南芥ATIPTs基因序列设计特异性引物,采用PCR方法从黄瓜S94基因组DNA中克隆出异戊烯基转移酶基因保守区片段;再通过对黄瓜基因组BAC文库的筛选,获得基因的全长序列,命名为cs-ipt,GenBank登录号HQ326777。经序列测定及分析表明,该基因编码序列全长1 005 bp,编码334个氨基酸,氨基酸序列中包含有ATP/GTP结合位点GATGTGKS。同源比对表明该基因的核苷酸序列和推导的氨基酸序列与其他植物的ipt基因都有较高的同源性,其中与杨树的同源性最高为65%。Primers were designed according to the ATIPTs genes sequence of Arabidopsls thaliana in GenBank.Partial sequences of the conserved region of isopentenyltransferases gene were cloned from cucumber S94 line by PCR methods,and then full-length sequences were obtained by filter the genomic BAC library from cucumber.Sequencing analysis showed that the gene（GenBank accession number HQ326777） was 1 005 bp length,encoded 334 amino acids,and included ATP/GTP binding site（GATGTGKS）.Homology analysis showed that the nucleotide and deduced amino acids were highly homologous to ipt gene from other species,specially the highest homology to ipt gene of Populus trichocarpa with the homology to 65%.According to the results of sequence analysis of cucumber ipt gene obtained in the study,we confirm that we have cloned ipt gene from Cucumis sativus,named cs-ipt.

关 键 词：黄瓜 IPT PCR BAC 克隆 

分 类 号：Q785[生物学—分子生物学]