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Integration and expression comparative analysis of heterologous and homologous dmrt1 expression vector in Medaka, Oryzias latipe
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1. Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture; Yellow Sea Fisheries Re-search Institute, Chinese Academy of Fishery Science, Qingdao 266071, China;2. College of Fisheries and Life Science, Shanghai Ocean Univers

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S917

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    Abstract:

    Fish are an ideal model for research on functional genes. Transgenic fish have many important applications in aquaculture. Recently, a number of researchers have evaluated the use of transgenic fish to improve the commercial value of a species by manipulating traits such as growth, sex, and disease resistance. In particular, the function of sex-determining genes has received significant attention. The current studies of gene function, which are related to sex differentiation, are largely confined to a few fresh water fish species. In contrast, little effort has been made to study gene function in marine fish because of logistical difficulties, such as the presence of a hard fertilized egg membrane, low survival rate, and/or long period of a generation. To address this gap, we studied the function of a homologous gene in a model marine fish species. A number of methods have been used to modify the fish genome, among which microinjection is the most commonly used approach. We constructed expression vectors by subcloning Cinto a pIRES- hrGFP-1α vector. in Medaka was induced by injecting these GFP constructs into the cytoplasm of fertilized Medaka embryos at the one cell stage. We then measured GFP expression and hatching rate. Additionally, we quantified the level of gene integration, expression, and the effect on expression. GFP was expressed at high levels 48 h after injection and the number of embryos expressing GFP decreased with the development of embryos. The hatching rate and integration efficiency of embryos injected with the homologous expression vector was significantly higher than those injected with the heterologous . Furthermore, in fishes with was down-regulated. Our research provides a foundation for the study of sex differentiation and contributes to our knowledge of sex-determining genes in marine fish.

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李亚亚,陈松林,林帆,王娜,刘洋,黄进强. 异源和同源dmrt1基因过表达载体在青鳉体内的整合和表达[J]. Jounal of Fishery Sciences of China, 2014,[volume_no](3):425-432

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  • Online: July 17,2015
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