大黄鱼肌肉组织cDNA全长文库的构建

bstract: A full length cDNA library from the skeletal muscle of Larimichthys crocea was constructed by using Creator Smart cDNA Library Construction Kit. This library reached 1.6×106 cfu/mL in apacity; the percentage of recombinant efficiency was as high as 96%. PCR results showed that the average size of inserts was larger than 1000 bp. 512 clones were randomly selected and sequenced. Of which, 430 expressed sequence tags (ESTs) were longer than 100 bp. 230 non-repetitive sequences were acquired, including 58 contigs and 172 singletons from these ESTs after initial assembly. Redundancy is 46.51%. By the retrieval, 35 ESTs are expected to be novel genes(E-value≤ 1.00E-10) because no sequence homology with any known sequences was found in GenBank batabases; 195 ESTs with known function were identified by Blastx. Energy-related genes of which had the highest abundance of ESTs, accounting for 20.00%, and protein synthesis, cytoskeleton and signal transduction followed by the proportion of 13.48%,12.17%, 10.00% respectively.The results are good enough for further screening and cloning of the specific expression genes in the skeletai muscle of Larimichthys crocea.