CONSTRUCTION OF CDNA EXPRESSION LIBRARY OF FLAMMULINA VELUTIPES

mRNA of Flammulina velutipes was extracted with Quick Prep mRNA Purification Kit, after synthesis of double-strand cDNA molecules using Time Saver cDNA Synthesis Kit, EcoRⅠ/NotⅠadaptors were added to the end of ds cDNA, phosphorylated by T4 polynucleoted kinase. Then the cDNA was ligated with λZAP Express Predigested Vector and packaged to transfect E.Coli XL1-Blue MRF? The results revealed that the tilter of the cDNA library was 4.0×106 pfu/ml, the amplified cDNA library is 2×1010 pfu/ml, the recombinant rate reached to 90%, and the length of inserts were between 0.3-3.5 kb. A candidate cDNA related with flammulin gene was screened by western blotting in situ.