论文部分内容阅读
为提高ｐＡＮ７—１质粒导入稻瘟病菌建立的遗传转化体系的转化率，进行了对转化效率影响因子的研究。稻瘟病菌转化溶液中使用５０ｍｍｏｌ／ＬＣａＣｌ２浓度，ＰＥＧ４０００４０％或ＰＥＧ８０００４０％多聚物，添加３０μｇ鲱鱼精ＤＮＡ，供体质粒ＤＮＡ呈直线状都有利转化频率的提高。其中鲱鱼精ＤＮＡ作用尤为显著，由于添加３０μｇ鲱鱼精ＤＮＡ，转化频率高达７１．６转化子／μｇＤＮＡ。４２℃热冲击对稻瘟病菌转化并非必需，转化溶液中添加受体菌２５３９ｗＤＮＡ后，反而引起转化频率的明显下降。
In order to improve the transformation rate of the genetic transformation system established by the introduction of pAN7-1 plasmid into Magnaporthe grisea, the factors affecting the transformation efficiency were studied. Magnaporthe grisea transformation solution using 50mmol / LCaCl2 concentration, PEG400040% or PEG800040% polymer, adding 30μg herring fish sperm DNA, donor plasmid DNA straight line favorable conversion frequency increase. Among them, herring sperm DNA was particularly effective. As the frequency of transformation was as high as 71.6 transformants / μg DNA due to the addition of 30 μg herring sperm DNA. Heat shock at 42 ℃ is not necessary for the transformation of Magnaporthe grisea. However, the addition of 2539wDNA to the transformation solution resulted in a significant decrease of the transformation frequency.