【摘要】 本项研究从新羽化的蜜蜂蜂王毒腺中提取了总mRNA，用逆转录的方法，合成了cDNA，并将其克隆到了噬菌体质粒λgtll的EcoRI位点，建立了melittin的cDNA文库。用PCR扩增技术从cDNA文库中产生了长度为87bp的melittin基因，并将其插入到高表达载体pBV220的EcoRI和pstI位点构成重组质粒PBM95，并转化到大肠杆菌JM101的感受态细胞中。经过在含氨苄青霉素的LB平板上对转化子进行筛选和对来自转化子中的重组质粒PBM95的酶切分析及melittin基因的测序，证明melit－tincDNA克隆成功。更多还原

【Abstract】 Total mRNA from venom glands of newly emerged queen bees was reversely transcribedinto cDNA and cloned into the EcoRI site of plasmid Agtll; cDNA library for bee venom was thus con-structed. PCR technique was used to preduce the melittin coding sequence from the cDNA library. A87 bp product was produced and inserted into the EcoRI and PstI sites of the high level expression vectorpBV220. Recombinant plasmid pBM95 was transformed into the competent cells of E.’coli JM101. Af-ter screening transformants on LB medium with ampicilin , structure of the recombinant plasmid pBM95from transformants was analyzed and melittin gene in pBM95 was sequenced. The cloned cDNA codingfor honey bee melittin was ohtained.更多还原