马蕾;李慧芬;彭忱晨;陈子柱;龙章富;

• 1:四川大学生命科学学院生物资源与生态环境教育部重点实验室

摘要(Abstract)：

[目的]研究蜡梅花香基因SAMT cDNA的克隆及表达。[方法]以蜡梅花瓣总RNA为模板进行RT-PCR,扩增得到与预期大小相同的PCR产物带,回收RT-PCR产物,将其与PMD18-T载体进行T-A克隆连接并导入大肠杆菌DH5α,经菌落PCR和双酶切鉴定,筛选带有目的基因的重组质粒并进行测序。[结果]测序证实成功克隆得到蜡梅花香基因SAMT cDNA的ORF片段,其长度为1 196 bp,编码380个氨基酸残基,与已报导的蜡梅SAMT(ABU88887)的同源性为99.2%。将SAMT基因亚克隆到原核表达载体PGEX4T-1中获得重组菌种命名为PGSAMT,用0.01 mol/L的IPTG进行诱导表达,经SDS-PAGE分析,SAMT的融合表达蛋白分子量约为66 kDa,与预期的26 kDa的GST带和42.3 kDa的蜡梅SAMT基因编码蛋白构成的融合蛋白大小接近。[结论]该研究成功克隆并表达蜡梅花香基因SAMT。

关键词(KeyWords)： 蜡梅;花香基因;cDNA;克隆;原核表达

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 马蕾;李慧芬;彭忱晨;陈子柱;龙章富;

Email:

DOI: 10.13989/j.cnki.0517-6611.2012.03.205

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