Identification of the nfa1 gene from N. gruberi transfected with the pEGFP-C2/nfa1UTR vector. (A) PCR for gDNA was performed with vector-specific (GFP F1) and nfa1-specific (nfa1 R3) primers. (B) The reverse transcription reaction for cDNA was performed with a vector-specific primer (VS1), and PCR was done with the GFP F1 and nfa1 R2 primers. Lanes 1, N. gruberi cultured at 33°C; lanes 2, N. gruberi transfected with the pEGFP-C2/nfa1UTR vector; lanes 3, positive control (plasmid DNA from E. coli DH5α transformed with the pEGFP-C2/nfa1UTR vector); lanes 4, plasmid DNA in lane 3 treated with 1 μg of DNase I; lanes M, One Kilobase Plus DNA ladder.