• Title/Summary/Keyword: Donor selection

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Development of Near-Isogenic Line of japonica Rice Cultivar Saenuri without Lipoxygenase-3 (새누리 벼 품종 배경 lipoxygenase-3 결핍 자포니카 근동질계통 개발)

  • Park, Hyun-Su;Lee, Keon-Mi;Kim, Ki-Young;Kim, Jeong-Ju;Shin, Woon-Cheol;Baek, Man-Kee;Kim, Choon-Song;Park, Seul-Gi;Lee, Chang-Min;Suh, Jung-Pil;Cho, Young-Chan
    • Korean Journal of Breeding Science
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    • v.51 no.3
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    • pp.190-200
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    • 2019
  • It is reported that the absence of lipoxygenase-3 (LOX-3) may contribute to a reduction in stale flavor after the storage of rice. To improve the quality of stored rice of the Korean japonica rice cultivar, we conducted a breeding program to develop near-isogenic rice without LOX-3 in the genetic background of Saenuri, a mega variety of Korea. In the first step of the breeding program, we used a donor parent of LOX-3 null, Daw Dam, and a recurrent japonica parent, Sindongjin, to develop HR27873-AC12 by backcross (BC1), color test for introgression of lox-3, and anther culture for rapid fixation. In the second step, we used the donor parent, HR27873-AC12, and the recurrent parent, Saenuri, to develop HR28896-31-3-1-1 by backcross (BC1), marker-assisted selection (MAS) for lox-3, and phenotypic selection (PS) for agronomic traits. Finally, in the third step, we developed HR30960-186-2-1-2-1 (Jeonju624), derived from a cross between Saenuri and HR28896-31-3-1-1, by MAS for lox-3 and PS with high selection pressure for agronomic characteristics. Jeonju624 was confirmed with the introgression of lox-3 by molecular marker. Jeonju624 was a mid-late maturing rice with similar agronomic characteristics to Saenuri, lodging tolerance with short culm, erect plant architecture, and resistance to bacterial blight and rice stripe virus. The yield components of Jeonju624 were mostly similar to Saenuri, except for the 1,000-grain weight of brown rice. The appearance of the grain of Jeonju624 was better than that of Saenuri, and the characteristics of cooked rice were similar to those of Saenuri. In the genetic background analysis using 406 KASP (Kompetitive Allele-Specific PCR) markers, Jeonju624 was confirmed to be the near-isogenic line (NIL) of Saenuri with a 95.8% recovery rate. Jeonju624 is the NIL of Saenuri without LOX-3, and overcomes the linkage drag of Daw Dam with similar agronomic characteristics and genetic background to Saenuri. Jeonju624 can be utilized as a practical cultivar to improve the quality of stored rice, breeding material for the introgression of lox-3, and genetic material to elucidate the effect of introgressed genes.

Characterization of Phylogenetic Incongruence among Protein Coding Genes of Vibrio Strains Pathogenic to Humans (인체 병원성 비브리오 균주간 유전자 계통의 불일치성 분석)

  • Zo, Young-Gun
    • Korean Journal of Microbiology
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    • v.49 no.4
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    • pp.383-390
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    • 2013
  • Lateral gene transfer (LGT) of genes from other bacteria into Vibrio cholerae is expectable because of the pronounced natural competence of the bacterium. In this study, quantitative aspects of LGT among the three species of Vibrio pathogenic to humans were characterized. Genome sequences of V. cholerae N16961, V. parahaemolyticus RIMD2210633, V. vulnificus CMCP6, and Escherichia coli K12 substrain MG1655 were analyzed to determine orthologous quartets of protein coding genes present in all four genomes. Phylogenetic analyses on the quartets were conducted to resolve vertical versus lateral patterns of gene polymorphisms based on congruence versus incongruence of phylogenetic trees. About 70% of the quartets could be resolved as either cohesive topology (75%) or LGT tree topologies (25%). The amount of LGT genes in Vibrio spp. appeared to be abnormally high for a genus and comparable to those of families. Patched distributions of LGT from different donors were observed on a chromosome. In the small chromosome of V. cholerae, physical linkages among LGT loci spanned half the length of the chromosome. Either accumulative selection for the donor alleles in LGT or presence of large-scale LGT events was hypothesized. These findings warrant further studies on the nature of donor-specificity of LGT alleles and its influence on evolution of Vibrio virulence to humans.

Generation and Selection of Promoter Trap Lines for the Investigation of Shoot Development in Arabidopsis (애기장대에 있어서 shoot 발달 연구를 위한 프로모터 trap 라인들의 제조 및 선별)

  • Lee Hwa-Mok;Park Hee-Yeon;Zulfugarov Ismayil S.;Lee Choon-Hwan;Moon Yong-Hwan
    • Journal of Life Science
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    • v.16 no.3 s.76
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    • pp.540-545
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    • 2006
  • T-DNA-mediated transformation is a common method for generating transgenic plants with insertional mutagenesis. In order to identify important genes involved in shoot development, a system of promoter trap insertional mutagenesis was employed in Arabidopsis thaliana. For this system, an efficient promoter trap vector, pFGL561 was developed. The pFGL561 includes a basta-resistant gene, an intron with multiple splicing donor and acceptor sites, and a promoter-less GFP reporter gene. Using floral-dipping method, we made total 300 $T_1$ promoter-trap lines which were screened for GFP expression. GFP signals in the $T_1$ plants were detected with high frequency, 26.7%, and the signals were reconfirmed in $T_2$ plants. To isolate the genes that are involved in shoot development, phenotypes were analyzed in $T_2$ plants of the 19 $T_1$ lines that had GFP signals in shoot apex, and 6 $T_1$ lines were selected that had abnormal shoot development. These lines will be very useful for the investigation of shoot development.

Liver Cut Method Using 4 Points for Hepatic Volumerty at MDCT Image (MDCT 영상에서 간 체적 계산을 위한 4 점 이용 간 분할 방법)

  • Seo, Jeong-Joo;Cho, Baik-Hwan;Park, Jong-Won
    • Journal of the Institute of Electronics Engineers of Korea SP
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    • v.47 no.1
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    • pp.17-24
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    • 2010
  • This paper proposed the method to separate a liver into left and right liver lobes for exact volumetry of the river graft at abdominal MDCT(Multi-Detector Computed Tomography) image before living donor liver transplantation. On the image of segmented liver, 4 points(the middle point of Inferior Vena Cava, a point of Middle Hepatic Vein, a point of Portal Vein, a middle point of gallbladder fossa) are selected. A liver is separated into left and right liver lobes on the basis of the 4 points. The volume and ratio of the river graft are estimated. The volume estimated using 4 points and the manual volume that radiologist processed and estimated are compared with the weight measured during surgery to support proof of the exact volumetry. After selection the 4 points, the time involved in separation a liver into left and right river lobe and volumetry of them is measured for confirmation that the algorithm can be used on real time during surgery. This study progressed to ensure donor's and recipient's safe who will undergo the liver transplantation.

Reproductive Biotechnologies for Improvement of Buffalo: The Current Status

  • Purohit, G.N.;Duggal, G.P.;Dadarwal, D.;Kumar, Dinesh;Yadav, R.C.;Vyas, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.7
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    • pp.1071-1086
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    • 2003
  • Reproductive biotechnologies continue to be developed for genetic improvement of both river and swamp buffalo. Although artificial insemination using frozen semen emerged some decades back, there are still considerable limitations. The major problem appears to be the lack of efficient methods for estrus detection and timely insemination. Controlled breeding experiments in the buffalo had been limited and similar to those applied in cattle. Studies on multiple ovulation and embryo transfer are essentially a replica of those in cattle, however with inherent problems such as lower number of primordial follicles on the buffalo ovary, poor fertility and seasonality of reproduction, lower population of antral follicles at all stages of the estrous cycle, poor endocrine status and a high incidence of deep atresia in ovarian follicles, the response in terms of transferable embryo recovery has remained low with 0.51 to 3.0 per donor and pregnancy rates between 15 to 30%. In vitro production of buffalo embryos is a valid alternative to recovery of embryos by superovulation. This aspect received considerable attention during the past decade, however the proportion of embryos that develops to the blastocyst stage is still around 25-30% and hence the in vitro culture procedures need substantial improvement. Embryo cryopreservation procedures for direct transfer post thaw need to be developed for bubaline embryos. Nuclear transfer and embryo cloning is a technique that has received attention in various species during recent years and can be of immense value in buffaloes as they have a low rate of embryo recoveries by both in vitro and in vivo procedures. Gender pre-selection, genome analysis, gene mapping and gene transfer are a few of the techniques that have been studied to a limited extent during recent years and are likely to be included in future studies on buffaloes. Very recently, reproductive biotechnologies have been applied to feral buffaloes as well, but the results obtained so far are modest. When fully exploited they can play an important role in the preservation of endangered species.

Screening of Sclerotinia Rot Resistant Korean Origin Perilla (Perilla frutescens) Germplasm Using a Detached Leaf Method

  • Lee, Ho-Sun;Afroz, Tania;Jeon, Young-Ah;Sung, Jung-Sook;Rhee, Ju-Hee;Aseefa, Awraris Derbie;Noh, Jaejong;Hwang, Aejin;Hur, On-Sook;Ro, Na-Young;Lee, Jae-Eun
    • Korean Journal of Plant Resources
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    • v.32 no.6
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    • pp.743-751
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    • 2019
  • Sclerotinia rot, caused by Sclerotinia sclerotiorum, is a devastating disease that poses a serious threat to perilla production in Korea. Identifying effective sources of resistance offers long term prospects for improving management of this disease. Screening disease resistant genetic resources is important for development of disease-resistant, new cultivars and conduct related research. In the present study, perilla germplasm were screened in vitro against S. sclerotiorum using detached leaf method. Among 544 perilla accessions, two were highly resistant (IT226504, IT226533), five were resistant (IT226561, IT226532, IT226526, IT226441, and IT226589), five were moderately resistant (IT226525, IT226640, IT226568, IT220624, and IT178655), 16 were moderately susceptible, 31 were susceptible, and 485 were highly susceptible. The resistant accessions in this study could serve as resistance donor in the breeding of Sclerotinia rot resistance or subjected to selection procedure of varietal development for direct use by breeders, farmers, researchers, and end consumers.

Current Status of Production of Transgenic Livestock by Genome Editing Technology (유전자 편집 기술에 의한 형질전환 가축의 생산 현황)

  • Park, Da Som;Kim, Soseob;Koo, Deog-Bon;Kang, Man-Jong
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.3
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    • pp.148-156
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    • 2019
  • The Transgenic livestock can be useful for the production of disease-resistant animals, pigs for xenotranplantation, animal bioreactor for therapeutic recombinant proteins and disease model animals. Previously, conventional methods without using artificial nuclease-dependent DNA cleavage system were used to produce such transgenic livestock, but their efficiency is known to be low. In the last decade, the development of artificial nucleases such as zinc-finger necleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regulatory interspaced short palindromic repeat (CRISPR)/Cas has led to more efficient production of knock-out and knock-in transgenic livestock. However, production of knock-in livestock is poor. In mouse, genetically modified mice are produced by coinjecting a pair of knock-in vector, which is a donor DNA, with a artificial nuclease in a pronuclear fertilized egg, but not in livestock. Gene targeting efficiency has been increased with the use of artificial nucleases, but the knock-in efficiency is still low in livestock. In many research now, somatic cell nuclear transfer (SCNT) methods used after selection of cell transfected with artificial nuclease for production of transgenic livestock. In particular, it is necessary to develop a system capable of producing transgenic livestock more efficiently by co-injection of artificial nuclease and knock-in vectors into fertilized eggs.

Enhanced Green Fluorescent Protein Gene under the Regulation of Human Oct4 Promoter as a Marker to Identify Reprogramming of Human Fibroblasts

  • Heo, Soon-Young;Ahn, Kwang-Sung;Kang, Jee-Hyun;Shim, Ho-Sup
    • Reproductive and Developmental Biology
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    • v.32 no.2
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    • pp.135-140
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    • 2008
  • Recent studies on nuclear transfer and induced pluripotent stem cells have demonstrated that differentiated somatic cells can be returned to the undifferentiated state by reversing their developmental process. These epigenetically reprogrammed somatic cells may again be differentiated into various cell types, and used for cell replacement therapies through autologous transplantation to treat many degenerative diseases. To date, however, reprogramming of somatic cells into undifferentiated cells has been extremely inefficient. Hence, reliable markers to identify the event of reprogramming would assist effective selection of reprogrammed cells. In this study, a transgene construct encoding enhanced green fluorescent protein (EGFP) under the regulation of human Oct4 promoter was developed as a reporter for the reprogramming of somatic cells. Microinjection of the transgene construct into pronuclei of fertilized mouse eggs resulted in the emission of green fluorescence, suggesting that the undifferentiated cytoplasmic environment provided by fertilized eggs induces the expression of EGFP. Next, the transgene construct was introduced into human embryonic fibroblasts, and the nuclei from these cells were transferred into enucleated porcine oocytes. Along with their in vitro development, nuclear transfer embryos emitted green fluorescence, suggesting the reprogramming of donor nuclei in nuclear transfer embryos. The results of the present study demonstrate that expression of the transgene under the regulation of human Oct4 promoter coincides with epigenetic reprogramming, and may be used as a convenient marker that non-invasively reflects reprogramming of somatic cells.

Analysis of 174 Consecutive Free Flaps (유리피판 이식술 174예의 분석)

  • Tark, Kwan Chul;Roh, Tai Suk
    • Archives of Reconstructive Microsurgery
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    • v.9 no.1
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    • pp.15-22
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    • 2000
  • One hundred & seventy four consecutive free-flap transfers were reviewed to analyze distribution of the type of reconstructions, kinds of donor flaps as well incidence of complications. The role of emergent exploration and the effect of preoperative wound conditions in flap survival were evaluated. Free flap transfer for head and neck reconstruction was most common as 93 cases, followed by for upper extremity of 30 cases, for lower extremity 30 cases, 18 penile reconstructions and for trunk & breast 3 cases. Nine flaps exhibited signs of ciruclatory insufficiency between 5 hours and 7 days. Three were managed conservatively with ultimate partial necrosis of the flaps. Eight flaps required return to the operating room. On exploration, early arterial occlusion was revealed in 1 flap, late arterial occlusion in 2 flaps, early venous occlusion in 1 flap, late venous thrombosis in 2 flaps, prolonged venous spasm in 1 and hematoma in 1 flap. The average time from the first abnormal examination to exploration was 2.6 hours. There were no false-positive explorations. Four free flaps failed in spite of the correction of the cause of circulatory compromise. The remaining 4 flaps were salvaged following the correction the casuse. Recipient vessel problems such as irradiation and infection were the most common cause of circulatory crisis. Among the eight flaps requiring return to the operating room, single vein was anastomosed in three flaps and two veins in the remaining five. In the totally failed four flaps only single vein was anastomosed in three cases. The results of this study demonstrate the efficacy of clinical monitoring and the role of early exploration. Precautious selection of recipient vessels and two vein anastomosis are recommended for safe and better prognosis.

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Selection of Various Free Flap Donor Sites in Palatomaxillary Reconstruction (구개상악재건을 위한 유리피판술에서 다양한 공여부의 선택)

  • Yoon, Do-Won;Min, Hee-Jun;Kim, Ji-Ye;Lee, Won-Jae;Chung, Seum;Chung, Yoon-Kyu
    • Archives of Reconstructive Microsurgery
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    • v.20 no.1
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    • pp.8-13
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    • 2011
  • Purpose: A palatal defect following maxillectomy can cause multiple problems like the rhinolalia, leakage of foods into the nasal cavity, and hypernasality. Use of a prosthetic is the preferred method for obturating a palate defect, but for rehabilitating palatal function, prosthetics have many shortcomings. In a small defect, local flap is a useful method, however, the size of flap which can be elevated is limited. In 12 cases of palatomaxillary defect, we used various microvascular free flaps in reconstructing the palate and obtained good functional results. Method: Between 1990 and 2004, 12 patients underwent free flap operation after head and neck cancer ablation, and were reviewed retrospectively. Among the 12 free flaps, 6 were latissimus dorsi myocutaneous flaps, 3 rectus abdominis myocutaneous flaps, and 3 radial forearm flaps. Result: All microvascular flap surgery was successful. Mean follow up time was 8 months and after the follow up time all patients reported satisfactory speech and swallowing. Wound dehiscence was observed in 4 cases, ptosis was in 1 case and fistula was in 1 case, however, rhinolalia, leakage of food, or swallowing difficultly was not reported in the 12 cases. Conclusion: We used various microvascular flaps for palatomaxillary reconstruction. For 3-dimensional flap needs, we used the latissimus dorsi myocutaneous flap to obtain enough volume for filling the defect. Two-dimensional flaps were designed with latissimus dorsi myocutaneous flap, rectus abdominis flap and radial forearm flap. For cases with palatal defect only, we used the radial forearm flap. In palatomaxillary reconstruction, we can choose various free flap techniques according to the number of skin paddles and flap volume needed.

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