[Expression analysis of a transformer gene in Daphnia pulex after RNAi].
In order to explore the importance of the transformer (tra) gene in reproductive mode switching in Daphnia pulex, we studied the effect of silencing of this gene using RNA interference (RNAi). We obtained Dptra dsRNA by constructing and using a dsRNA expression vector and transcription method in vitro. D. pulex individuals in different reproductive modes were treated by soaking in a solution of Dptra dsRNA. We then assayed the expression of the endogenous Dptra mRNA after RNAi treatment using RT-PCR and obtained the suppression ratio. Expression of the tra gene in the RNAi groups was down-regulated compared with the controls after 16 h (p < 0.05). We also analyzed the effect of RNAi on the expression of the TRA protein using Western blot, which showed that the expression level of the TRA protein was reduced after RNAi treatment. Our experimental results showed that soaking of D. pulex adults in tra-specific dsRNA transcribed in vitro can specifically reduce the level of tra mRNA and also reduce the expression of the TRA protein, demonstrating effective in vivo silencing of the tra gene.
School of Marine Sciences, Ningbo University, Ningbo, 315211 China.
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CY
Guo
School of Marine Sciences, Ningbo University, Ningbo, 315211 China.
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P
Chen
School of Marine Sciences, Ningbo University, Ningbo, 315211 China.
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Zhang
School of Marine Sciences, Ningbo University, Ningbo, 315211 China.
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Ning
School of Marine Sciences, Ningbo University, Ningbo, 315211 China.
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Wang
School of Marine Sciences, Ningbo University, Ningbo, 315211 China.|wangdanli@nbu.edu.cn.
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Wang
School of Life Science, East China Normal University, Shanghai, 200062 China.
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Zhao
D000818:Animals; D060829:Arthropod Proteins / Q000096:biosynthesis* / Q000235:genetics; D003621:Daphnia / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology*
D060829:Arthropod Proteins; D009687:Nuclear Proteins; D034741:RNA, Small Interfering
Western blotting; cladocerans; parthogenesis; reproductive transformation