Construction of cDNA libraries from cultures of Armillaria mellea

Abstract

1. 1. Conditions were established for growth of mycelial cultures of Armillaria mellea such that the production of its lysine-specific proteinase was maximized. Proteinase synthesis was confirmed by immunoprecipitation. 2. 2. Mycelia grown under these same conditions were used as a source of RNA and this RNA was translatable in a wheat germ translation system to produce proteins with M r in the range < 10,000–> 90,000 3. 3. Double-stranded cDNA was prepared and was inserted into the EcoR1 site of λgt10 and λgt11 using an adaptor ligation strategy. Packaging of these materials yielded large cDNA libraries. That from λgt10 contained 2.9 sx 10 6 pfu/ml with 70% recombinants whereas that from λgt11 contained 2.2 × 10 6 pfu/ml with 60% recombinants.