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A Converging Strategy for the Generation of a Virtually Sequenced cDNA Library from Unreferenced Pacific Oysters
Published on: June 13, 2019
Generation and analysis of expressed sequence tags from a cDNA library of Moniezia expansa
Wen Juan Zhao1, Hui Zhang, Xinwen Bo
1Genetic Engineering Laboratory, The Breed & Biotechnology Key Laboratory of Sheep in XinJiang, Bingtuan 832000, PR China.
Abstract:
The Moniezia expansa is a parasite of sheep that causes diarrhea and fleshless leading to stockbreeding losses. A genomic resource for large-scale molecular studies in this parasite is lacking. To study the gene expression including development, digestion and reproduction organs of M. expansa, a cDNA library had been constructed and expressed sequence tags (ESTs) had been analyzed, which were helpful for the development of a powerful microarray platform which are used to analyze gene expression in this species. cDNAs are useful resources in annotating genes and providing functional analysis of genes. In this study, a cDNA library from adult cestode of M. expansa was created and 2642 ESTs from 5'-ends of the cDNA clones representing 1081 unigenes were obtained. Cluster analysis of these ESTs allowed identification of 1081 unique sequences containing 351 contigs and 730 singletons. BLASTX searches identified 780 significant (E-value<10(-5)) hits and further Gene Ontology (GO) analysis was used to annotate these genes. All the EST sequences were deposited under dbEST in GenBank (GenBank: FE905224-FE905315, FE942104-FE942773, FE969189-FE969190, FF677548-FF677734, FF848124-FF848253). Although we only obtained 1081 unigenes, the set of ESTs identified represents a significant proportion of the M. expansa and provides molecular resource for the development of microarrays for gene expression studies concerning development, metabolism and reproduction.
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