双孢蘑菇不同交配型同核不育单孢的基因组文库构建

Genomic Library Construction of Monokaryons with Different Mating Type of Agaricus bisporus

对24个双孢蘑菇不育单孢进行两两杂交配对,以菌株9608为基准,获得了2种不同交配型的不育单孢菌株A+:9601、9608、9609、9612、AgQG841-1、AgLH830-4、AgLH830-6、AgQL8125-5;A-:9602、9603、9604、9607、Ag2k811-1、M7206-1、M7206-2、M7206-11、M7206-13。将这2种菌株进行DNA提取,并构建DNA文库,对文库进行酶切验证,用SalⅠ酶切,可切出20、14、9kb等3个片段,其中20、9kb为λ载体的左右臂,插入片段为14kb,与预期相符,说明文库的质量较高。

In this study,twenty-two Agaricus bisporus monokaryons strains were pairwisely paired.Based on strain 9608,two different mating types of monokaryons were obtained.A + strains:9601,9608,9609,9612,AgQG841-1,AgLH830-4,AgLH830-6,AgQL8125-5.A-:9602,9603,9604,9607,Ag2k811-1,M7206-1,M7206-2,M7206-11,M7206-13.The genomic DNA of two strains were extracted and DNA library were constructed.The library was verified by restriction enzyme digestion with SalI.The 20 kb,14kb and 9kb 3fragments were cut out,of which 20 kb and 9kb were the inserted into the left and right arms of lambda vector and the fragment of 14 kb,was the insertion genomic DNA fragments.Enzyme digestion result was in line with expectations,indicating the higher quality of the library.