• Applied Biological Chemistry
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• v.36 no.3
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• pp.208-214
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• 1993

To investigate the gene rearrangement via short repeated sequences in chloroplast DNA, the pattern of heterologous gene clusters containing the 151 bp repeated sequence with the development of plastid was compared in rice and the homologous gene clusters from various plant sources were searched for comparative analysis. Southern blot analysis of rice DNA using rp12 gene containing 151 bp repeated sequence as a probe showed the presence of heterologous gene clusters. Such heterologous gene clusters varied with the development of plastid. Also it was observed that the heterologous gene clusters were observed in all of the rice cultivars used in this work. Finally the comparative analysis of DNA sequence of the homologous gene clusters from various plants showed the evolutionary gene rearragngement via short repeated sequence among plants. These results suggest the possible relationship between the plastid development and gene rearrangement through short repeated sequences.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2023.04a
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• pp.19-19
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• 2023

제비꽃속 34분류군의 61개체를 대상으로 엽록체 DNA 게놈 특이 유전자의 특징을 알아보고자 하였다. 61개체의 엽록체 게놈 전체 길이는 155,535~158,940 bp 로 모두 전형적인 사분할 구조였다. 지역별로는 LSC 지역이 84,826~87,250 bp, SSC 지역이 16,338~18,654 bp, 그리고 IR 지역이 26,029~27,192 bp 였다. 유전자 개수는 131개로 84개 protein coding-gene, 37개 tRNA 유전자, 8개 rRNA유전자, 그리고 2개의 유사유전자인 𝜓rps19, 𝜓ycf1으로 구성되어 있었다. LSC/IRa 경계에 위치한 rps19 유전자 길이는 279 bp로 모든 분류군에서 동일하였으며, 𝜓rps19의 길이는 다양했으나 유전자 개수에는 영향을 미치지 않았다. SSC/IRb 경계에 위치한 ycf1 유전자 길이는 약 5,600 bp 였으나, V. japonica (MZ151699) 1개체에서는 다른 종에 비해 약 1,000 bp 위치에서 발생한 점돌연변이로 인해 종결 코돈이 나타나는 특징을 보였다. 한편 13분류군의 23개체에서는 𝜓ycf1의 길이가 650 bp 정도 짧은 것을 확인하였는데, 이 종류들은 원예종인 V. tricolor (ON262802) 이외에는 모두 줄기가 없는 분류군들로 IR 지역의 확장과 SSC 지역의 수축에 의한 것으로 판단된다. ndhF는 대체로 SSC 지역에 위치하나, V. inconspicua (MZ065354), V. mongolica (MW802534, ON548135), V. yunnanfuensis (MW802541) 등 4개체에서는 IRa/SSC 경계에 위치하면서 유사유전자가 발생하였고, 그 결과 다른 제비꽃 종류에 비해 유전자 개수가 132개로 차이를 보였다. 또한, V. collina (OP271831), V. mirabilis (MH256000), V. tricolor (ON262802) 등 3분류군에서는 SSC 지역이 inversion 되어 엽록체 이성질체가 존재함을 확인하였다. 이상의 결과를 종합하면, 제비 꽃속 엽록체 게놈 61개체의 ycf1, 𝜓ycf1, ndhF, 𝜓ndhF 등은 유전자 길이와 개수 등에 차이를 보이는 것으로 나타났으며, 제비꽃속에서도 엽록체 이성질체가 존재함을 확인할 수 있었다.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2021.04a
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• pp.22-22
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• 2021

큰졸방제비꽃(Viola kusanoana)의 엽록체 DNA 염기서열을 밝히고자 차세대염기서열분석법(NGS)을 이용하여 분석하였다. 재료는 경상북도 울릉군 나리분지에 자생하는 개체의 잎을 사용하였다. 염기서열 분석결과, 총 길이는 158,644 bp 였고, GC함량은 36.3%로 분석되었다. 구간별로는 LSC (Large single copy)지역이 86,999 bp (GC content: 33.9%)였고 SSC (Small single copy)지역은 17,439 bp (GC content: 29.9%)으로 분석되었으며 IR (Invertied repeats)지역은 27,103 bp (GC content: 42.2%)로 확인되었다. 유전자는 protein coding gene 77개, tRNA gene 30개, rRNA 4개 등 총 111개로 이는 선행 연구된 제비꽃속 8개 분류군과 유전자의 순서와 방향이 모두 일치하였다. 이를 통해 제비꽃속의 엽록체 게놈의 유전자는 상당히 보존되어 있음을 확인하였다.

• Journal of Life Science
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• v.20 no.4
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• pp.481-486
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• 2010

The rates of synonymous and nonsynonymous nucleotide substitutions were studied for sequences of nuclear and chloroplast genes in Sorbus aucuparia. Results suggested that DNA evolution in this species had taken place, on average, at a slower rate in the chloroplast genes than in the nuclear genes: a rate variation pattern similar to those observed in eudicot plants. Within the nucleus, the synonymous substitution rates (Ks) (2.45-2.60) were two-fold higher than nonsynonymous substitution rates (Ka) (1.15-1.30). More notably, the values of Ks (1.20-1.26) were about six-fold higher than those of Ka (0.26-0.42) within the chloroplast genome. Ka/Ks ratios for nuclear and chloroplast genes of S. aucuparia had mean values of 0.178 and 0.056, respectively. A Ka/Ks ratio < 1 indicated negative (purifying) selection. The chloroplast genes had a lower effective number of codons (ENC) values (22.4-32.2) than those of nuclear genes (35.8-38.7). The analysis of the G+C content indicated that the chloroplast genes in this investigation had a higher preference for synonymous codons ending with A and T (G+C content range, 28.4-29.1%) where there was a slight bias toward codons ending with G+C (63.2-64.2%) in the nuclear genome.

• Journal of Ginseng Research
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• v.16 no.1
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• pp.80-89
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• 1992

• Journal of Plant Biotechnology
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• v.37 no.3
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• pp.275-282
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• 2010

Chloroplast transformation in higher plants offers many attractive advantages over nuclear transformation, including a high-level accumulation of foreign proteins, multi-gene expression in single transformation event via transgene stacking in operons and no position effect due to site-specific integration of transgenes by homologous recombination. Most importantly, chloroplast transgenic plants are eco-friendly because their transgenes are maternally inheritance in most crop plants. However, chloroplast transformation system has limited success in crops alike nuclear transformation. In the past two decades, great progress has been made to overcome the limitations of chloroplast transformation, thus expending chloroplast bioreactor to several important crops including soybean, carrot, lettuce, and oilseed. Therefore, it has become possible that chloroplast transformation of crops can be used not only for the improvement of agronomic traits, but also for the production of vaccines and high valuable therapeutic proteins in pharmaceutical industry.

• Journal of Plant Biotechnology
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• v.34 no.3
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• pp.271-275
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• 2007

In the chloroplast transformation process, a chloroplast containing transformed chloroplast genome copies should be selected over wild-type chloroplasts on selection medium. It is more effective for a cell to become homoplasmic if the cell contains smaller number of chloroplasts. Therefore, to reduce the number of chloroplasts in mesophyll cells in tobacco, we overexpressed FtsZ to generate transgenic plants, of which mesophyll cell contained a few enlarged chloroplasts contrast to a wild-type mesophyll cell containing approximately 100 chloroplasts. It was demonstrated that transgenic leaf tissues comprising cells with a few enlarged chloroplasts gave rise to approximately 40% higher frequency of chloroplast-transformed adventitious shoots.

• Proceedings of the Botanical Society of Korea Conference
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• 1990.09a
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• pp.155-172
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• 1990

• Korean Journal of Plant Taxonomy
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• v.45 no.3
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• pp.213-220
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• 2015

To elucidate the origin and patterns of establishment of insular plants on Ulleungdo Island, maternally inherited chloroplast DNA, which is useful for tracing seed movements, was used. Fagus multinervis, an endemic species that dominated broadleaf deciduous forests on Ulleungdo Island, is an excellent model for such a study. To understand the diversity and spatial distribution of the chloroplast haplotypes of F. multinervis, nucleotide sequences of the psbA-trnH region were determined from 144 individuals sampled throughout the island. Results of a phylogenetic analysis of the region with close relatives of F. multinervis suggest that F. multinervis is sister to a clade of F. japonica and F. engleriana. No haplotype variation was found within F. multinervis. This remarkably low cpDNA haplotype diversity is in contrast to the findings of previous allozyme studies of F. multinervis populations that showed high genetic diversity on Ulleungdo Island. Repeated colonization during the early stage of establishment via birds that migrated from a source area where the F. multinervis cpDNA haplotype was geographically structured may have resulted in the observed pattern of haplotype diversity. Alternatively, long-distance dispersal of seeds of the progenitor of F. multinervis via birds or typhoons to Ulleungdo may have been a single event, whereas the immigration of pollen from the mainland likely occurred frequently. Comparative phylogeographic studies of other species endemic to Ulleungdo Island and their close relatives on the neighboring mainland are necessary for a more complete understanding of the evolution of the island's native species.

• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.42-48
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• 2011

Chloroplast genetic engineering of higher plants offers several unique advantages compared with nuclear genome transformation, such as high levels of transgene expression, a lack of position effect due to site-specific transgene integration by homologous recombination, multigene engineering in a single transformation event and reducing risks of gene flow via pollen due to maternal inheritance. We established a reproducible chloroplast transformation system of potato using a tobacco specific plastid transformation vector, pCtVG (trnI-Prrn-aadA-mgfp-TpsbA-trnA). Stable transgene integration into chloroplast genomes and the homoplasmic state of the transgenome were confirmed by PCR and Southern blot analyses. Northern, immunoblot analysis, and GFP fluorescence imaging revealed high expression and accumulation of GFP in the plastids of potato leaves. This system would provide new opportunities for genetic improvement and mass production of value added foreign proteins in this crop.