Isolation and Transcriptome Analysis of Ophiocordyceps sinensis Fungus

H. sinensis L0106 was isolated from wild O. sinensis samples that were gingerly collected from Qinghai-Tibet Plateau in Qinghai Province. H. sinensis L0106 has been deposited at the China Center for Type Culture Collection (Wuhan, China) with accession number CCTCC M2011278. Subsequently, the transcriptome of H. sinensis was sequenced and the reads obtained were further assembled into Unigenes, and then BLASTx alignment (e < 0.00001) between protein databases and Unigenes was performed [21 (link)]. E. coli JM109 was selected as host for plasmid pMD18-T transformation (Invitrogen, Carlsbad, CA, USA), and E. coli BL21 (DE3) (Invitrogen) was selected as the host for expression of pET28a (Invitrogen). Lysogeny broth (LB) medium (10 g/L tryptone, 5 g/L yeast extract, and 10 g/L NaCl) at 37°C with shaking (200 rpm) was used for the growth of E. coli transformants. Ampicillin, kanamycin, isopropyl-β-D-thiogalactopyranoside (IPTG), adenosine, vernine, cytidine, uridine, and thymidine standard substances were purchased from Sigma Chemical Co. (St. Louis, MO, USA).