title: Construction of differentially expressed genes library of bighead carp (Aristichthys nobilis) exposed to microcystin-lr using ssh and expression profile of related genes.
abstract: Microcystins (MCs) are hepatotoxic cyclic heptapeptides produced by cyanobacteria (blue-green algae). There are more than 70 MCs variants of which the most common and widely studied is MC-LR. We screened the hepatocellular differentially expressed genes against MC-LR in the bighead carp (Aristichthys nobilis). Suppression subtractive hybridization was used to construct the forward subtracted and reverse subtracted cDNA libraries, and one hundred and thirty two positive clones (seventy one in forward library and sixty one in reverse library) were randomly selected and sequenced. Finally, fifty five reliable sequences from the forward subtracted library were used in a homology search by BLASTn and BLASTx, as were 57 reliable sequences from the reverse subtracted library. Furthermore, eight analyzed sequences from the forward subtracted cDNA library and seven from the reverse subtracted library were found to be non-homologous sequences. The screening identified genes induced by MC-LR in both libraries that are involved in various processes, such as energy metabolism, immunity, and apoptosis. Some are cytoskeleton- and transportation-related genes, while signal transduction-related genes were also found. Significant genes, such as the apoptosis-related gene p53 and the proto-oncogene c-myc, are involved in inhibition of the MC-LR response in the reverse subtracted library. In addition, several immune-related genes, which play an important role in antioxidation and detoxification of MC-LR, were characterized and identified in both of the subtracted libraries. The study provides the basic data to further identify the genes and molecular mechanism of detoxification of microcystins.
authors.affiliation: College of Fisheries and Life Sciences, Shanghai Ocean University, 999, Hucheng Huan Road, Shanghai, PR China.
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Cui
authors.forename: Kaiyue
authors.initials: K
authors.lastname: Zhang
authors.forename: Xiancheng
authors.initials: X
authors.lastname: Qu
authors.forename: Qigen
authors.initials: Q
authors.lastname: Liu
mesh_terms: D000818:Animals; D001483:Base Sequence; D002347:Carps / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D019295:Computational Biology; D017931:DNA Primers / Q000235:genetics; D020869:Gene Expression Profiling / Q000662:veterinary*; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000276:immunology; D015723:Gene Library; D008099:Liver / Q000378:metabolism*; D008387:Marine Toxins; D052998:Microcystins / Q000633:toxicity*; D008969:Molecular Sequence Data; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D017422:Sequence Analysis, DNA / Q000662:veterinary
chemical_list: D017931:DNA Primers; D008387:Marine Toxins; D052998:Microcystins; C057862:cyanoginosin LR