Construction of cDNA Library and Preliminary Screening of Immune-Related Genes in Cyclina sinensis

To investigate the molecular mechanism of immune of Cyclina sinensis, a cDNA library infected by Vibrio anguillarum was constructed based on SMART technique using Construction Kit (Clontech). The result of the test showed that the primary constructed cDNA library had a cloning capacity of 1.12×10⁶ and the length of the insert fragments was above 500bp, therefore, the quality of the library could meet the requirements. By conducting high-throughput sequencing on white clones randomly selected from the library and informatics analysis of the sequences, 1113 expressed sequence tags (ESTs) out of which 126 contigs were obtained. Through the Blastx on NCBI non-redundant protein database comparation, the total number of annotated sequences was 271 (64.5%), including 46 immune related sequences. These results could serve as the foundation of screening and cloning the full-length functional immune genes of Cyclina sinensis.