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Construction of cDNA expression library from adult Fasciola gigantica and preliminary analysis of its expressed sequence tags



Construction of cDNA expression library from adult Fasciola gigantica and preliminary analysis of its expressed sequence tags



Acta Zoologica Sinica ober; 51(5): 879-883



To determine the latent vaccine molecules that resist Fasciola gigantica disease and to provide resources for further study, a high-quality Fasciola gigantica cDNA library was constructed into [lambda] TriplEx2 plasmid using the SMART technique, the result showed that the unamplified library capacity was 1.08 X 106 PFU/ml, the library recombination rate was 96.6%, the titer of the amplified library was 2.41 X 109 PFU/ml and the average inserted cDNA fragment was about 1 000 bp. Then 40 clones were selected to sequence from the BM25.8 plasmids, 32 sequences obtained from the plasmids proved useful after editing using biosoftware. After analysing using the BLASTx and BLASTn programs, 9 Expressed Sequence Tags (ESTs) were known-genes, and 16 ESTs whose alignment values were very low or did notmatch known sequences were considered unknown-genes. The 9 ESTs represent cystein protease, eggshell protein and calcium-binding protein. The 16 unknown ESTs included some signal conductive genes, protein translative genes and immune stimulative genes and so on. The Fasciola gigantica cDNA expression library has established the base for research into its bio-elements and mechanism ( Acta Zoologica Sinica 51 (5) : 879 - 883, 2005).

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