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포인세티아로부터의 dihydroflavonal-4-reductase(DFR) 유전자의 분리 및 분석 : Euphorbia pulcherrima
구자정,박영두 경희대학교 생명자원과학연구원 2004 硏究論文集 Vol.23 No.-
본 연구는 포인세티아 포염색을 결정하는 화색관련유전자 dihydroflavonal-4-reductase(DFR)을 분리하고 그 기능을 분석하기 위해 수행되었다. DFR specific primer를 제작하여 RT-PCR을 통해 498 bp의 단편을 얻었으며 염기서열 분석을 통해 Gerbera hybrid와 98%의 상동성을 갖는 DFR 유전자임을 확인하였다. 기능 분석을 위하여 분리된 DFR 유전자를 CxVMV promoter에 의해 조절되는 식물 발현 벡터 Agrobacterium을 이용하여 담배(Nicotiana tobaccum cv. Havana SR1)에 형질전환하였다. 형질 전환 식물체는 잎 절편체를 재조합 벡터를 포함한 A. nanefaciens LBA440에 접종한 후 1, 2차 배지에서 신초 및 뿌리를 유도함으로서 확보할수 있었다. 1차 PCR에 의해 marker 유전자의 전이가 확인된 개체는 온실에서 재배하여 자가수분 및 채종을 하였으며 genomic DNA를 추출하여 DFR 유전자가 담배 genome 내로 전이되었음을 PCR로 다시 확인하였다. 채종된 종자는 후대 유전 분석을 위해 kanamycin이 첨가된 파종배지에서 선발중에 있다. DFR specific primer was designed to isolate dihydroflavonal-4-reductase (DFR) from Euphorbia pulcherrima cv. Feedom Red. The DFR gene fragment was isolated through RT-PCR with a set of primers designed based on homologous coding region present in DFR genes from divergent sources. To analyze the function of DFR gene, an antisense-orineted DFR gene fragment amplified by RT-PCR was cloned into a binary vector PILTAB 357 for plant transformation. This construct was introduced into Agrobacterium tumefaciens strain LBA4404 and used for transformation into the Nicotiana tabaccum cv. Havana SRI. The transformation of the antisense-orineted DFR gene into tobacco was confirmed by PCR.
구자갑,김운학,최정호,유영화,정양규 한경대학교 2004 論文集 Vol.26 No.1
This paper study on soil properties and earth pressure at the domestic multi layer. Reliable predictions of the movement of earth retaining structures and the ground adjacent to the braced walls in urban excavation are often difficult due to many variable factors. Also, Unpredictable behavior of ground and retaining structure due to variation of the above factors may cause considerable damage to the adjacent structures, and cost many of human lives as a results of retaining wall failure. The earth pressure at the retaining walls was measured at total 13 numbers sites and Peck's empirical earth pressures adapted at the design stage. The geotechnical properties by this research was proposed as a preliminary design guide line in urban excavation where controling ground movement and safety assurance of adjacent structure. The earth pressure of rock masses were found to be 70.14% of Peck's earth pressure and the earth pressure of soil were found to be 44.4% of Peck's earth pressure. It is indicated that the design earth pressures of a retaining wall is
Ku, Ja-Jung,Kim, Yong-Yul The Plant Resources Society of Korea 2008 한국자원식물학회지 Vol.21 No.3
Gene-expression analysis is increasingly important in biological research, with real-time reverse PCR (RTPCR) becoming the method of choice for high-throughput and accurate expression profiling of selected genes. However, this technique requires important preliminary work for standardizing and optimizing the many parameters involved in the analysis. Plant stress studies are more and more based on gene expression. The analysis of gene expression requires sensitive and reproducible measurements for specific mRNA sequence. Several genes are regulated in response to abitoic stresses, such as salinity, and their gene products function in stress response and tolerance. The design of the primers and TaqMan probes for real-time PCR assays were carried out using the Primer $Express^{TM}$ software 3.0. The PCR efficiency was estimated through the linear regression of the dilution curve. To understand the expression pattern of various genes under salt stressed condition, we have developed a unique public resource of 9 stress-related genes in poplar. In this study, real-time RT-PCR was used to quantify the transcript level of 10 genes (9 stress-related genes and 1 house keeping gene) that could play a role in adaptation of Populus davidiana. Real-time RT-PCR analyses exhibited different expression ratios of related genes. The data obtained showed that determination of mRNA levels could constitute a new approach to study the stress response of P. davidiana after adaptation during growth in salinity condition.
A New Soybean Cultivar with Lodging Resistance and Adaptability for Mechanized Harvesting, "Shingi"
Jung Kyung Moon,Keum Yong Park,Hong Tae Yun,Yeong Ho Lee,Min Jung Seo,Sun Lim Kim,Yong Hwan Ryu,Suk Ha Lee,Ja Hwan Ku,Jae Hwan Roh,Jung Soo Park,Ik Jae Kim,Soo Kyeong KimMoon 한국육종학회 2005 한국육종학회지 Vol.37 No.2
Methylation Pattern Analysis of Double-Transformed Tobacco Plants with Antisense-Oriented MET1 Gene
Ja Jung Ku,Chae Wan Lim,Young-Doo Park 한국원예학회 2006 Horticulture, Environment, and Biotechnology Vol.47 No.6
DNA methylation in plants is related to a number of epigenetic phenomena. Heavy methylation of cytosine residues plays an important role in gene expression, and significant differences in cytosine methylation levels have been observed among various tissue types, which can be explained as one of the regulatory mechanisms during development and differentiation. Tobacco (Nicotiana tabaccum cv. Havana SR1) was transformed with Agrobacterium tumefaciens LBA4404 containing a rescue cloning vector, pRCV2. The T-DNA tags were isolated by rescue cloning. These intact T-DNA tags, obtained by rescue cloning from transgenic lines, were fully sequenced to analyze the actual insertion site of T-DNA in tobacco. In the case of mutant line single transformant (ST) #5-1 and 2, blast search of plant flanking DNA were shown homology with N. tabaccum RENT family: tCUP promoter. Flanking plant DNA of ST #2-3 and #8-0 clones showed significant aligriments to H2 trans-silencing gene and enoyl-acyl carrier protein reductase, respectively. Transgenic plants transformed with pRCV2 were retransformed with A. tumefaciens EHA105 carrying pFTM vector. Restriction enzyme digestion patterns of RENT repetitive sequence were analyzed using DNA extracted from the STs and the double transformants (DTs) in order to determine whether transcriptional gene silencing (TGS) of MET1 affected DNA methylation. As a result, STs with dsMET1 decreased methylation in all DTS. This result means that DT-MET1 transformants causes hypomethylation of the STs, presumably due to TGS of MET1.