文献类型：期刊文献

中文题名：根癌农杆菌介导载体pBI121转化少根根霉条件的研究

英文题名：Conditions of Agrobacterium tumefaciens mediated transformation of Rhizopus arrhizus with pBI121 plasmid

作者：赵有玺[1];冀颐之[1];欧阳艳华[1];龚平[1]

第一作者：赵有玺

机构：[1]北京联合大学生物化学工程学院

第一机构：北京联合大学生物化学工程学院

年份：2014

卷号：39

期号：7

起止页码：46-49

中文期刊名：食品科技

外文期刊名：Food Science and Technology

收录：北大核心:【北大核心2011】；

基金：北京市优秀人才培养项目(2012D005022000007);启明星大学生科技创新项目(201411417SJ085);北京联合大学人才强校计划人才项目

语种：中文

中文关键词：pBI121;根癌农杆菌介导的转化;少根根霉

外文关键词：pBI121; Agrobacterium tumefaciens-mediated transformation; Rhizopus arrhizus

摘要：考察了脂肪酶生产菌株少根根霉N1023 G418的敏感性,结果表明,G418对少根根霉最小抑制浓度为500μg/mL。研究了不同条件对载体pBI121转化少根根霉的影响,结果表明,以根癌农杆菌LBA4404为介导菌,少根根霉N1023为受体菌株,G418为筛选标记的最佳转化条件为:乙酰丁香酮(AS)300μmol/L、根癌农杆菌的初始浓度OD600为0.6、共培养时间2 d。在此条件下pBI121对少根根霉的转化率达到112 cfu/106 cfu孢子。转化菌株经过5次连续培养后仍然稳定。通过上述研究,建立了根癌农杆菌介导载体pBI121转化少根根霉的方法。这是首次成功建立少根根霉的转化方法,为进一步的研究奠定了基础。
Rhizopus arrhizus was a strain which can produce lipase. The sensitivity of Rhizopus arrhizus to G418 was investgated. The minimal inhibitory concentration （MIC） for G418 of Rhizopus arrhizus was 500 μg/mL. Agrobacterium tumefaciens-mediated transformation of Rhizopus arrhizus with pBI121 plasmid was studied and the optimized parameters were investigated. The results show that the optimized parameters were： acetosyringone （AS） concentration 300 μmol/L, the concentration of Agrobacterium tumefaciens OD600 0.6, co-culture time 2 d. Under the optimized conditions, the transforming efficiency of Rhizopus arrhizus with pBI121 reached 112 transformants/106 spores. The transformants were still stable after the successive cultivation for 5 times. In study, we firstly established the method of transformation of R. arrhizus. This lay the foundation for further research.