CONSTRUCTION OF cDNA LIBRARY WITH APOSTICHOPUS JAPONICUS CULTURED IN THE SOUTH AND SEQUENCE ANALYSIS OF ITS TROPOMYOSIN GENE

Based on the construction technology of non-normalized cDNA library, the cDNA libraries of Apostichopus japonicus cultured in the south was constructed. This library reached 2.2×107 in capacity; the percentage of recombination was 95.8%. PCR results showed that the average size of inserts was larger than 750bp. A total of 185 successful sequencing reactions yielded. In addition, 136 non-repetitive sequences were formed including 40 contigs and 96 singletons from these ESTs after initial assembly. The tropomyosin gene of A. japonicus, which was 1112bp long, its ORF encoding 284 amino acids with a predicted molecular weight of 33.27kDa and a theoretical isoelectric point of 4.56. The amino acid sequence was compared with Strongylocentrotus purpuratus, Euphausia superba, Portunus sanguinolentus, Branchiostoma belcheri, Eriocheir sinensis, and Bombyx mori; and high similarities of 62%—46% were revealed.