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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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白木香倍半萜合酶基因asss4的克隆、原核表达与功能鉴定
, PP. 1724-1729
Keywords: 白木香,倍半萜合酶基因,原核表达,基因功能
Abstract:
通过rt-pcr方法从伤害处理的白木香树木质部中克隆得到沉香倍半萜合酶4(sesquiterpenesynthase4,asss4)基因的全长开放读码框,该基因全长读码框为1698bp,编码包含565个氨基酸的蛋白质.将asss4基因与原核表达载体pet28a相连构建重组载体pet28a-asss4,把重组载体转入大肠杆菌bl21(de3)plyss中,用iptg诱导重组菌,经sds-page电泳分析,获得分子质量约为64kd的重组asss4蛋白.利用镍凝胶亲和色谱法获得纯度较高的asss4蛋白,以法尼基焦磷酸(fpp)为底物进行蛋白酶促反应,共催化产生5种倍半萜类成分:cyclohexane,1-ethenyl-1-methyl-2,4-bis(1-methylethenyl)-、β-elemene、α-guaiene、α-caryophyllene和δ-guaiene.本研究首次证实了白木香倍半萜合酶asss4基因的功能,为揭示伤害诱导沉香倍半萜形成的分子机制奠定了理论基础.
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