神秘果素基因合成及其液泡积累表达载体DC-miraculin的构建  

Synthesis of miraculin gene and construction of its plant expression vector DC-miraculin with miraculin accumulated in vacuole

作　　者：王新卫[1] 章镇[1] 朱月林[1] 高志红[1] 乔玉山[1] 

机构地区：[1]南京农业大学园艺学院,江苏南京210095

出　　处：《南京农业大学学报》2010年第6期38-42,共5页Journal of Nanjing Agricultural University

基　　金：国家科技重大专项重点课题项目(2009ZX08004-011B)

摘　　要：在不依赖神秘果(Synsepalum dulcificum Daniell)植物资源的条件下,根据已报道的神秘果素基因序列合成24条寡核苷酸引物,通过重叠延伸PCR法合成了神秘果素全基因。为使神秘果素导向液泡积累,利用菜豆蛋白C末端四肽的液泡定位功能,在人工合成基因的3'末端加入了编码液泡定位短肽的核苷酸序列。将该基因序列插入植物表达载体YH4215,成功构建了神秘果素植物双元表达载体,命名为DC-miraculin。载体上的神秘果素基因由双CaMV35S启动子控制,转基因植物的报告基因为gusA,抗性筛选标记为潮霉素抗性基因hpt。The miraculin gene was synthesized from 24 chemical synthesized oligo primers based on the reported nucleotide sequence of miraculin by overlap extension PCR method,though the lack of gene template. In order to direct the miraculin to accumulate in vacuole,the nucleotide sequence of C-terminal tetrapeptide of phaseolin with sufficient information for vacuolar sorting was fused to 3＇ terminal of the miraculin gene,and the modified gene was inserted into the plant expressing vector YH4215,constructing a new vector designated DC-miraculin for expressing miraculin in plant. Miraculin gene in this vector was under the control of double CaMV35S promoter,and gusA and hpt were the reporter gene in transgenic plant and hygromycin resistance gene as selected marker,respectively.

关 键 词：神秘果素 基因合成 液泡 载体构建 

分 类 号：Q812[生物学—生物工程]