一种用于PCR扩增的丝状真菌DNA快速提取方法

Rapid Extraction of Filamentous Fungal DNA for PCR Amplification

摘要:

丝状真菌在工业、农业、医药以及基础生物学研究中具有重要作用。利用遗传转化技术对丝状真菌进行菌株改良和基因功能分析, 也越来越受到重视。然而, 丝状真菌DNA提取方法繁琐、费时, 难以满足利用PCR技术高通量筛选转化子的需要。本文以曲霉菌为例建立了一种快速提取丝状真菌DNA的实验方法, 微波处理置于10 × TE buffer中的菌丝即可得到DNA。RAPD试验和PCR扩增证明, 该方法提取的DNA能够达到PCR扩增的要求。研究结果为高通量快速筛选丝状真菌转化子奠定了基础。

Abstract:

Filamentous fungi play an important role in industry, agriculture, medicine and biological fundamental research. More and more attention has been paid to fungal breeding and gene functional analysis via genetic transformation technology. However, the extraction of fungal DNA was labor intensive and time-consuming, which makes it difficult to perform high-throughput screening of transformants through PCR technology. In this paper, a rapid DNA extraction method was established for filamentous fungi (take Aspergillus for example). DNA was easily obtained by exposing the fungal mycelia in 10 × TE buffer to microwave. RAPD and PCR experiments demonstrated that the extracted DNA satisfied the requirements of PCR amplification. These results provided a foundation for the high-throughput screening of filamentous fungal transformants.