...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Microbiological Research >An efficient Agrobacterium-mediated transformation method for the edible mushroom Hypsizygus marmoreus
【24h】

An efficient Agrobacterium-mediated transformation method for the edible mushroom Hypsizygus marmoreus

机译：一种有效的农杆菌介导的食用菌玛格氏菌转化方法

获取原文
获取原文并翻译 | 示例
   

获取外文期刊封面封底 >>

       
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Hypsizygus marmoreus is one of the major edible mushrooms in East Asia. As no efficient transformation method, the molecular and genetics studies were hindered. The glyceraldehyde-3-phosphate dehydrogenase (GPD) gene of H. marmoreus was isolated and its promoter was used to drive the hygromycin B phosphotransferase (HPH) and enhanced green fluorescent protein (EGFP) in H. marmoreus. Agrobacterium tumefaciens-mediated transformation (ATMT) was successfully applied in H. marmoreus. The transformation parameters were optimized, and it was found that co-cultivation of bacteria with protoplast at a ratio of 1000:1 at a temperature of 26 degrees C in medium containing 0.3 mM acetosyringone resulted in the highest transformation efficiency for Agrobacterium strain. Besides, three plasmids, each carrying a different promoter (from H. marmoreus, Ganoderma lucidum and Lentinula edodes) driving the expression of an antibiotic resistance marker, were also tested. The construct carrying the H. marmoreus gpd promoter produced more transformants than other constructs. Our analysis showed that over 85% of the transformants tested remained mitotically stable even after five successive rounds of subculturing. Putative transformants were analyzed for the presence of hph gene by PCR and Southern blot. Meanwhile, the expression of EGFP in H. marmoreus transformants was detected by fluorescence imaging. This ATMT system increases the transformation efficiency of H. marmoreus and may represent a useful tool for molecular genetic studies in this mushroom species
机译：马齿more（Hypsizygus marmoreus）是东亚主要的可食用蘑菇之一。由于没有有效的转化方法，分子和遗传学研究受到阻碍。分离了马氏链球菌甘油三磷酸甘油醛脱氢酶（GPD）基因，并用其启动子驱动马氏链球菌中的潮霉素B磷酸转移酶（HPH）和增强型绿色荧光蛋白（EGFP）。根癌农杆菌介导的转化（ATMT）已成功地应用于Marmoreus。优化了转化参数，发现在含有0.3 mM乙酰丁香酮的培养基中，在26摄氏度的温度下，以1000：1的比例与原生质体细菌共培养导致了农杆菌菌株的最高转化效率。此外，还测试了三个质粒，每个质粒带有驱动抗生素抗性标记表达的不同启动子（来自H. marmoreus，灵芝和香菇）。携带H. marmoreus gpd启动子的构建体比其他构建体产生更多的转化体。我们的分析表明，即使在连续五轮继代培养之后，超过85％的测试转化子仍保持有丝分裂稳定。通过PCR和Southern印迹分析推定的转化体中hph基因的存在。同时，通过荧光成像检测了EGFP在H.marmoreus转化株中的表达。该ATMT系统提高了H. marmoreus的转化效率，可能代表该蘑菇种进行分子遗传学研究的有用工具

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号