不结球白菜晚抽薹BcFLC3基因的克隆及表达分析  

Cloning and Expression Analysis of Late Bolting BcFLC3 Gene from Brassica campestris ssp. chinensis

作　　者：荣子龙[1] 侯喜林[1] 史公军[1] 肖栋[1] 王倩[1] 张琳[1] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室农业部南方蔬菜遗传改良重点开放实验室,南京210095

出　　处：《西北植物学报》2010年第12期2373-2378,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：江苏省自然科学基金(BK2009311);江苏省科技支撑计划(BE2008377)

摘　　要：根据大白菜BcFLC3基因(GenBank登录号AY036890.1)保守域序列设计引物,扩增不结球白菜晚抽薹BcFLC3基因的核心片段,结合RACE技术获得该基因1 017 bp的全长cDNA序列。序列分析结果表明,该cDNA包含完整开放阅读框,编码197个氨基酸的蛋白质,其分子量为21.62 kD,等电点9.36。荧光定量PCR分析表明,不结球白菜经4℃低温处理后,BcFLC3基因在叶片中的表达量抽薹前明显高于抽薹后;低温处理后BcFLC3基因在不同部位的表达量存在明显差异,表达量由高到低依次为叶、茎、花蕾、花和根。Southern杂交结果表明,Bc-FLC3基因在不结球白菜基因组中为多拷贝。Brassica campestris ssp.pekinensis was used as a test material.A pair of degenerating primers was designed according to the conserved regions of BcFLC3 gene（AY036890.1） from B.campestris ssp.pekinensis and a cDNA fragment was isolated from B.campestris ssp.chinensis using RT-PCR.Full-length cDNA with 1 017 bp was acquired by RACE methods.Its ORF（open read frame） encoded a polypeptide of 197 amino acids.The putative protein of this gene had an isoelectric point of 9.36 and a calculated molecular weight of 2.162×10^4.Real-time PCR analysis showed expression level of the BcFLC3 gene in unbolting plant＇s leaves was higher than that of bolting plant.After vernalization treatment,the expression variation in different tissues of bolting plant was detected.The expression level from high to low were leaf,stem,bud,flower and root.The Southern blot showed that BcFLC3 had multi-copy in B.campestris ssp.chinensis.

关 键 词：不结球白菜 FLC基因 荧光定量PCR SOUTHERN杂交 

分 类 号：Q785[生物学—分子生物学]