배추 유래 전신발현 프로모터의 동정
Identification of Constitutive Promoters Derived from Brassica rapa
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초록
Promoters are essential regulatory elements for efficiently expressing a gene of interest in a target tissue of a organism. Therefore, the identification of a suitable promoter is important in plant biotechnology. In this study, four promoters were selected and identified to be constitutively or tissue-specifically expressed in Brassica rapa bacterial artificial chromosome (BAC) clones using the results of transcriptomic analysis of Brassica rapa and open-source database information on Arabidopsis thaliana. The 2 kb region of the 5′ upstream was isolated from the Brassica rapa genomic DNA for each promoter. The four promoters were then fused to β-glucuronidase (gus) and green fluorescent protein (gfp) genes, and the recombinant transgenes were introduced into Arabidopsis. As a result of histochemical GUS staining, GFP fluorescence and RT-PCR, the gus gene was observed to be expressed constitutively in all tissues using all four promoters. A GUS activity assay using fluorescent 4-MUG revealed that the BR11 promoter showed similar activity to the CaMV35S promoter, while the BR4, BR15, and BR16 promoters showed 1.5, 4, and 18-fold higher activities than the CaMV35S promoter, respectively. These results indicate that these four promoters could be used to incorporate useful genes with enhanced function into crops of interest.
Abstract
Promoters are essential regulatory elements for efficiently expressing a gene of interest in a target tissue of a organism. Therefore, the identification of a suitable promoter is important in plant biotechnology. In this study, four promoters were selected and identified to be constitutively or tissue-specifically expressed in Brassica rapa bacterial artificial chromosome (BAC) clones using the results of transcriptomic analysis of Brassica rapa and open-source database information on Arabidopsis thaliana. The 2 kb region of the 5′ upstream was isolated from the Brassica rapa genomic DNA for each promoter. The four promoters were then fused to β-glucuronidase (gus) and green fluorescent protein (gfp) genes, and the recombinant transgenes were introduced into Arabidopsis. As a result of histochemical GUS staining, GFP fluorescence and RT-PCR, the gus gene was observed to be expressed constitutively in all tissues using all four promoters. A GUS activity assay using fluorescent 4-MUG revealed that the BR11 promoter showed similar activity to the CaMV35S promoter, while the BR4, BR15, and BR16 promoters showed 1.5, 4, and 18-fold higher activities than the CaMV35S promoter, respectively. These results indicate that these four promoters could be used to incorporate useful genes with enhanced function into crops of interest.
참고문헌 (42)
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