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      저자 : 현정오 ( Suk Gyu Son (검색결과 2 건)
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        Agrobacterium tumefaciens vector 를 이용한 양황철의 형질전환 (

        손석규,현정오 ( Suk Gyu Son,Jung Oh Hyun ) 한국산림과학회 1998 한국산림과학회지 Vol.87 No.2

        This study was conducted to find the optimum transformation condition using Agrobacterium harboring promoterless GUS gene. The optimal medium for shoot induction from leaves of Populus nigra×P. maximowiczii was MS medium supplemented with 0.1㎎/ℓ NAA, 0.5㎎/ℓ BAP(94% regeneration frequency and 11.5 average number of shoot) According to the test using pBI121, the concentration of antibiotics for selection marker gene was 100㎎/ℓ kanamycin or 60㎎/ℓ geneticin in the SIM(shoot inducing medium) 3. Two weeks later, callus was induced in the SIM 3 and this callus grew up to 0.5-1cm shoots after 6 weeks in the new SIM 3. And the treatment with methylation inhibitor(5-azacytidine) led to a dramatic increase in foreign gene expression rate from 5.7% to 26.7% . The vector systems showed. different transformation efficiencies based on the fluorometric and histochemical GUS assay. In this study the vector systems used for transformation seemed to affect transformation frequency, in which pEHA101 yielded more transformants(35.9%) than LBA4404/pBI121 did(5.7%). This result indicated that pEHA101 was effective to insert the promoterless foreign gene into a poplar genome.

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        Inverse PCR 기법을 이용한 양황철 DNA 의 Regulatory Region 의 탐색

        손석규,현정오 ( Suk Gyu Son,Jung Oh Hyun ) 한국산림과학회 1998 한국산림과학회지 Vol.87 No.3

        This research was conducted to identify plant regulatory regions by gene tagging method. A promoterless GUS coding sequence was introduced to Populus nigra × P. maximowiczii via Agrobacterium strains(LBA4404/EHA101), and putative transgenic poplars were selected by culturing on medium containing G418(60㎎/ℓ) and by GUS assay. Among them one positive plant was to amplify the native sequences flanking to the introduced GUS gene in plant genome by inverse PCR method and from this 730 by DNA product was obtained. After subcloning and sequencing, it has 88% homology to the Eucalyptus gunnii CAD(cinnamyl alcohol dehydrogenase) gene. The GUS gene fused with the putative promoter reinserted into poplar leaves by particle bombardment method to test the funtional promoter activity. Upon staining with X-gluc, many blue sprots appeared on the leaf segments bombarded by the chimeric gene 2-3 days, thus the isolated DNA fragment contain some possible coding region as well as a putative regulatory sequences of poplar CAD gene.

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