摘要
柞蚕抗菌肽具有广谱杀菌功能，人工合成抗菌肽Ｄ基因（１２２ｂｐ）与含α－因子及前导肽序列的穿梭质粒ｐＣＬＷＡ２重组，克隆于酵母ＡＢ１０３。在缺亮氨酸的ＹＥ培养基中筛选Ｌｅｕ＋阳性的转化子。转化子菌株培养扩增后提取重组质粒与抗菌肽基因片段探针作点杂交为阳性，电泳后插入含抗菌肽Ｄ基因１２８ｂｐ的区带。在转基因酵母的发酵液中分离上清液，浓缩，对抗菌肽敏感的指示菌Ｅ．ｃｏｌｉＫ１２Ｄ３１有明显抑菌效应。证实人工合成柞蚕抗菌肽Ｄ基因在酵母中表达。
Antibacterial peptide developed from haemolyrnph of Chinese oaksilkworm , An theraea pernyi,had broad spectyum inhibition on bacteria. The antibacterial peptide Dgene (122bp)was recombined with a shuttle plasmid pCL WA2 containing a promotor of alpha-factor. Transfered the recombinant plasmid into yeast AB103.A medium YE without leucine was used to select the transformant(Leu+).Extracted the recombinant plasmid from transformant yeast and made dot blot with a probe of antibacterial peptide gene. The resuIt showed a 128bp DNA fragment containing antibacterial peptide D gene was inserted into yeast and identified by PAG electrophoresis. The fermentive product of transformant yeast N 15 had a strong inhibition on E. coli Ki2D31. That indicted a gene of antibacterial peptide D from Chinese oak silkworm was expressed in yeast.
出处
《蚕业科学》
CAS
CSCD
1994年第2期95-99,共5页
ACTA SERICOLOGICA SINICA
基金
广东省自然科学基金
关键词
柞蚕
抗菌肽D基因
酵母
Chinese oak silkworm Antibacterial peptide gene Yeast
