水稻ERECTA基因组DNA的克隆及植物表达载体构建  

Cloning of Rice ERECTA Genomic DNA and Construction of Plant Expression Vector

作　　者：韩同凯[1,2] 王盈盈 林红珍[3] 陈翠霞[1] 谢先芝 

机构地区：[1]山东农业大学农学院,山东泰安271018 [2]山东省水稻研究所/山东省水稻工程技术研究中心,山东济南250100 [3]山东省农业科学院农产品研究所,山东济南250100

出　　处：《山东农业科学》2013年第6期4-10,共7页Shandong Agricultural Sciences

基　　金：国家自然科学基金(30870192);山东省自然科学杰出青年基金(2009JQ11)资助

摘　　要：根据水稻ERECTA的cDNA序列(GenBank中的登陆号:Os06g0203800),利用NCBI的Blast工具找到OsERECTA所对应的基因组DNA序列(gERECTA),根据该序列设计2对特异引物,对gERECTA进行分段PCR扩增,并对前后两部分片段进行克隆、测序和拼接。本研究得到的gERECTA序列为8 901 bp,其中包括约1.7 kb的5'区域和约300 bp的3'区域。将拼接起来的gERECTA序列插入到pCAMBIA1390载体,经酶切和测序鉴定,证明重组表达质粒中带有gERECTA基因片段,表明gERECTA植物表达载体pCAMBIA1390-gERECTA已成功构建。Based on the reported OsERECTA cDNA sequence in rice （GenBank accession number： Os06g0203800）, its corresponding genomic DNA sequence （gERECTA） was researched by the Blast tool of NCBI. Two pairs of primers were designed to isolate the upstream and downstream segments separately, and then the two segments were joined together to form an intact gERECTA gene and subcloned into an expression vector pCAMBIA1390. The full length of the cloned gERECTA was 8 901 bp, including 1.7 kb of 5＇and 300 bp of 3＇regions. The recombinant plasmid pCAMBIA1390 -gERECTA was identified by double digestion and sequencing, which suggested that the plant expression vector pCAMBIA1390 -gERECTA was successfully constructed.

关 键 词：水稻 OsERECTA基因 克隆 植物表达载体 

分 类 号：Q78[生物学—分子生物学]