Cloning and characterization of shrimp IRF

Shrimp total RNA was extracted using RNeasy Mine Kit (Qiagen, Germany) and reverse transcribed into cDNA using a PrimeScript^TM RT reagent kit (TaKaRa, Japan). A partial cDNA sequence homologous to mammalian IRFs was retrieved from the sequenced L. vanname transcriptome data56 (link), and primers IRF-5RACE1 and IRF-3RACE1 were then designed to receive the 3′ and 5′ ends of IRF cDNA sequences by rapid amplification of cDNA ends (RACE). PCR program was set as described before57 (link). The PCR products were used as templates with the primers IRF-5RACE2 and IRF-3RACE2 for the secondary PCR, with the PCR conditions the same as above. The PCR products were then cloned into the pMD-19T vector (TaKaRa, Japan) and sequenced. The sequences were analyzed and deposited in the NCBI GenBank (GenBank accession no. KM277954).