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Construction and analysis of suppressive subtractive hybridization library from Perna viridis induced by triazophos |
SUN Wei, ZHANG Lin-bao, HU Ying, CAI Wei-gui, JIA Xiao-ping |
Key Laboratory of Fishery Ecology Environment, Guangdong Province, Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China |
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Abstract To study the reproductive toxicity and molecular mechanism of triazophos to marine shellfishes, a suppressive subtractive hybridization (SSH) cDNA library of gonads from female Perna viridis by actue triazophos (1mg/L) stress was constructed for the first time. A total of 117expressed sequence tags (ESTs) from the forward and reverse subtraction library exhibited homology with genes of know functions, 12ESTs related to hypothetical protein, and the remaining 195 ESTs had no significant matches in the GenBank database. Among the 117 annotated sequences,the most abundant upregulated transcripts identified in the forward SSH libraries were associated with cell cycle regulation, sex steroid biosynthesis, DNA repairing and respiratory processes, while genes encoding proteins of DNA replication, transcription and translation regulation, cytoskeleton were found in reverse SSH libraries. Altogether, the results showed that different pathways (reproduction control, respiratory processes and stress adaptation) of mussel metabolism were changed in both forward and reverse directions after exposed to triazophos. Several reproduction-related transcripts (cyclin, cyclin-dependent kinase, zygotic DNA replication licensing factor, sperm-specific protein, estrogen sulfotransferase and vitelline envelope zona pellucida domain) identified in forward library might be the key genes involved in reproductive disturbance of triazophos, and provided the basis for the study of reproductive toxicity of organophosphorus pesticide.
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Received: 02 April 2016
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