摘要
研究比较 3种方法提取白僵菌DNA的效果并给予改进。结果表明 ,在无液氮条件下 ,可用氯化苄促使细胞破裂 ,释放DNA ,但此法效果不很稳定 ,容易造成DNA降解断裂或收率很低。SDS和CTAB都能很好地分离析出DNA ,后者的收率较高 ,可达 >50 μg/10 0mg菌丝。CsCl对清除多糖和蛋白质残余有很好的作用。用改进的方法得到高纯度、高收率的白僵菌DNA ,并可用于酶切或PCR扩增。
The effect of 3 methods of extracting DNA from Beauveria bassiana were compared and improved. The results of the experiments showed that SDS (sodium dodecyl sulfate) and CTAB (cetly triethyl ammonium bromidewere) both are effective for isolate DNA from the fungus. A higher yield of DNA could be obtained by CTAB, which get to 50 μg/100 mg fresh mycelia. Benzyl Chloride could break cells and made them release DNA without being ground in liquid nitrogen. But the protocol tended to cause lower yield and more DNA digestion. Polysacchrides from fungal cell was removed with CsCl. High quality and quantity of DNA from the fungus could be provided by the modified procedure with CTAB, which was practiccal for isolating DAN from a large number of fungal samples. The extracted DNA was suitable for RAPD PCR and RFLP PCR analysis.
基金
农业部农作物病虫草害生物防治资源研究与利用重点实验室资助
关键词
DNA提取
白僵菌
真菌
DNA extraction
Beauveria bassiana
Fungus
