植物甜蛋白monellin基因的合成和克隆

SYHTHESIS AND CLONING OF cDNA ENCODING THE SWEET PLANT PROTEIN MONELLIN

根据已报道的单链 monellin基因序列 ,设计合成 5段核苷酸序列 ,利用这些片段末端的碱基互补序列经链延伸、PCR扩增合成双链 DNA,克隆于质粒载体 p U C18的 Bam HI、Sal I位点之间 ,经限制酶切分析 ,序列分析等方法证实获得了 m onellin基因的克隆 ,与报道的 monellin基因的序列完全符合 ,为下一步构建高效表达质粒 ,并将monellin基因转入植物并获得高含量的 m

Five specific primers were designed according to genomic sequence of monellin reported in literature.The DNA was synthesized by chain extension,followed by PCR amplification.The synthesized DNA was cloned into plasmid pUC18 in E.coli JM 109.The recombinant plasmid was screened and further identified by restriction enzyme analysis and nucleotide sequence analysis.The results showed that the sequence of the cloning is same as that of monellin reported in literature.