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虾肝肠胞虫(Enterocytozoon hepatopenaei, EHP)基因组不同提取方法的比较
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Abstract:
为探寻最佳EHP基因组提取方法，本研究采用传统酚/氯仿抽提法、改良酚/氯仿抽提法、几丁质酶法、商品化海洋动物DNA提取试剂盒及商品化植物DNA提取试剂盒共5种方法分别对等量的EHP进行基因组提取。提取的基因组浓度由高到低依次为：商品化海洋动物DNA提取试剂盒(18.825 ng/μL) > 几丁质酶法(17.17 ng/μL) > 改良酚/氯仿抽提法(15.06 ng/μL) > 传统酚/氯仿抽提法(10.8 ng/μL) > 商品化植物DNA提取试剂盒(2.175 ng/μL)；提取的基因组完整性由高到低依次为：商品化海洋动物DNA提取试剂盒 > 改良酚/氯仿抽提法 > 商品化植物DNA提取试剂盒 > 传统酚/氯仿抽提法 > 几丁质酶法。由上述结论可知，采用商品化海洋动物DNA提取试剂盒提取的EHP基因组浓度最高，且完整性最好，是5种方法中最佳的提取方法。
In order to explore the most suitable extraction method of the Enterocytozoon hepatopenaei (EHP) genome, five methods were used to extract genomes of equal amounts of EHP, including traditional and improved phenol/chloroform extraction, enzymatic hydrolysis of chitin, marine animals DNA kit and plant genomic DNA kit. Results showed that the effect on the genomic concentration was in descending order of marine animals DNA kit (18.825 ng/μL), enzymatic hydrolysis of chitin (17.17 ng/μL), improved phenol/chloroform extraction (15.06 ng/μL), traditional phenol/chloroform extraction (10.8 ng/μL), plant genomic DNA kit (10.8 ng/μL); the effect on the genome integrity was in descending order of marine animals DNA kit, improved phenol/chloroform extraction, plant genomic DNA kit, traditional phenol/chloroform extraction, enzymatic hydrolysis of chitin. Due to the highest extraction concentration and the best integrity, this study confirms that the method of marine animals DNA kit was the most suitable extraction method for EHP genome.
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