Journal of Aquaculture (한국양식학회지)

The Korean Society of Fisheries and Aquatic Science (한국양식학회)
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Analysis of Fish Expression Vectors for Construction of Two MARs Expression Vector System in Fish Cell Line

  • Lim, Hak-Seob (Department of Bioteclmology and Bioengineering Pukyong National University) ;
  • Park, Jin-Young (Department of Bioteclmology and Bioengineering Pukyong National University) ;
  • Hwnag, Jee-Hwang (Department of Bioteclmology and Bioengineering Pukyong National University) ;
  • Kim, Moo-Sang (Department of Bioteclmology and Bioengineering Pukyong National University) ;
  • Lee, Hyung-Ho (Department of Bioteclmology and Bioengineering Pukyong National University)
  • Published : 2000.03.01
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Abstract

In previously study we isolated several fish matrix attachment regions (MARs) capable of replicating the plasmid by itself. In this study we construct a fish expression vector pBaEGFP(+) containing mud loach ${\beta}$-actin promoter EGFP as reporter gene and SV40 signal. To analyze the effects of the fish expression vector respectively. The fish ARS containing constructs pBaEGFP(+)-ARSs were transfected cells with pBaEGFP(+)-ARS101 and pBaEGFP(+)-ARS223 reduced 10 days to 25 days and then was constant to 30 days after transfection while that of the control vector without ARS element was basal level. The intensity of both constructs showed about 30fold of the intensity compared with the control vector on 30days after transfection individually .E. coli back-transformation analysis shows that pBaEGFP(+)-ARS223 and pBaEGFP(+)-ARS905 maintain in episomal state at least 30 days after transfection. The result indicates that both may be able to replicate the vector in BF-2 cell. Therefore the matrix-attached ARSs enhancing expression of the reporter gene might be useful as a component o the expression vector for transgenic studies.

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References

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