• Title/Summary/Keyword: LBA

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Acquisition of Thermotolerance in the Transgenic Plants with BcHSP17.6 cDNA (BcHSP17.6 cDNA의 도입에 의한 형질전환된 식물의 내열성 획득)

  • Ki Yong Kim;Min Sup Chung;Jin Ki Jo
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.17 no.4
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    • pp.379-386
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    • 1997
  • Recombinant plasmid, pBKH4, containing NPT II and P35S-BcHSP17.6 was constructed by ligation of Bum H I -digested pBKSl-l and BcHSP 17.6 (thermotolerance gene) 6om pBLH4. The tobacco leaf disc was cocultivated with transformed Agmbacterium tumefaciens bearing pBKH4 for 24 hours and transformed shoots were selected on MS-n/B medium containing $100\;{\mu\textrm{g}}/ml$ of kanamycin. Heat-killing temperature of Nicotima tabacum was $50^{\circ}$ for >15min, and transformed tobacco plants with BcHSP17.6 cDNA exhibited thermotolerance at the heat-killing temperature. The transgenic plants were analyzed by Southern blot hybridization with the probe of ${\alpha}^{_32}P$ labelled BcHSP17.6 cDNA. Transcription and expression level of BcHSP17.6 cDNA were also continued by Northern blot analysis and Ouchterlony double immunodiffusion assay. In this study, we suggest that the BcHSP17.6 cDNA introduced to tobacco plant is related to thenuoto-lerance and 17.6-kD LMW HSP acts as a protector from heat damage in plants.

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Transgenic Plants Expressing an Antisense RNA of ALl-Gene from Tomato Golden Mosaic Virus(TGMV) (Tomato Golden Mosaic Virus(TGMV) AL1 -gene의 antisense RNA 발현 형질 전환 식물체)

  • 임성렬
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.147-152
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    • 1998
  • AL1-gene, necessary for the replication of the genome of a gemini virus TGMV, was inserted in the opposite direction to the promoter CaMV35S resulting in the construction of a plant transformation binary vector pAR35-2. The vector pAR35-2 contains the chimeric gene cassette involving the duplicated promoter CaMV35S, opposite direction of AL1-gene fusioned with hygromycin resistant gene, and the gene cassette of the neomycin phosphotransferase II gene. The plasmid was transferred to tobacco and tomato plants by leaf disk infection via Agrobacterium. The transgenic plants were selected and grown on the MS-agar medium containing kanamycin and hygromycin. The shoots induced from the calli were regenerated to the whole transgenic plants. The antisense AL1-gene was detected in the genomic DNA isolated from the leaves by using the PCR mediated Southern blot analysis. The expression of the antisense AL1-gene was also observed using the RT-PCR mediated Southern blot analysis. The observation of chloroplasts in guard cell pair indicated that the transgenic tomato plants were diploid.

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Efficient Transformation Method of Soybean Using Meristematic Tissues of Germinating Seeds (발아종자의 분열조직을 이용한 효율적인 콩 형질전환 방법)

  • Kim, Yul-Ho;Park, Hyang-Mi;Choi, Man-Soo;Sohn, Soo-In;Shin, Dong-Bum;Lee, Jang-Yong
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.278-285
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    • 2008
  • An efficient transformation method for soybean [Glycine max (L.) Merr.] using meristematic tissues of germinating seeds has been established. The embryonic axes were excised from germinating seeds of Korean soybean cultivar, Iksannamulkong and 0.5-2 cm long segment containing meristematic tissues were prepared by cutting hypocotyl region. The explants were inoculated with Agrobacterium tumefaciens strain LBA4404 harboring a binary vector with the bar gene as a selectable marker gene and a ${\beta}-glucuronidase$ (GUSINT) reporter gene, and then co-cultured for 7 days on co-cultivation medium (CCM). The meristematic tissues were cultured on shoot induction medium (SIMP6) supplemented with 0.4 mg/l $N_6-benzylaminopurine$ (BAP) and 0.1 mg/l indolebutyric acid (IBA) in the presence of 6 mg/l L-phosphinotricin (PPT) for 2 weeks and the surviving explants were transferred to shoot elongation medium (SEMP6). Transformation was confirmed by Southern blot analysis and the transformation efficiencies ranged from 1.48 to 2.07%. The new modified transformation method was successfully implemented for obtaining several transgenic lines with SMV-CP gene. It is expected that this method could efficiently be used for the transformation of recalcitrant soybean cultivars.

Introduction of CAX1 into 'Hongro' Apple via Agrobacterium tumefaciens (CAX1 유전자가 도입된 사과 '홍로' 형질전환체)

  • Kim, Jeong-Hee;Shin, Il Sheob;Cho, Kang-Hee;Kim, Se Hee;Kim, Dae-Hyun;Hwang, Jeong Hwan
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.534-539
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    • 2010
  • 'Hongro' is early-mid maturing cultivar with good quality like 'Tsugaru' and it has not preharvest drop. The CAX1 gene was introduced into Korean apple cultivar 'Hongro' by Agrobacterium tumefaciens LBA4404 harboring pBI121 to obtain transgenic apple with enhanced Ca level. The CAX1 gene playing the role of $H^+/Ca^{2+}$ transporter from Arabidopsis thaliana increases Ca concentration in several plants. Regenerated transgenic lines were confirmed by polymerase chain reaction (PCR) analysis and Southern blot analysis of genomic DNA for the existence of CAX1 gene. Southern blot analysis of 'Hongro' transformants showed that two putative transgenic lines were integrated with CAX1 gene in genomic DNA. The CAX1 comparative expression levels of two transgenic lines were higher than that of non-transformant evaluated by comparative quantification analysis using a real-time PCR. These two lines were multiplied in vitro, and micro-grafted on apple rootstocks 'M.9' in the isolated greenhouse. Since two years after micro-grafting, the fruits came into bearing. Compared to Ca level of the non-transgenic 'Hongro', that of the CAX1 transgenic 'Hongro' in the flesh and leaves was higher.

The Apple Rootstock Transgenic M.26 (Malus pumila) with Enhanced Rooting Ability (발근력이 향상된 사과 대목 M.26 형질전환체)

  • Kim, Jeong-Hee;Kwon, Soon-Il;Shin, Il Sheob;Cho, Kang-Hee;Heo, Seong;Kim, Hyun Ran
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.482-487
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    • 2009
  • The apple rootstock M.26 (Malus pumila) is very popular apple rootstock with semi-dwarf habit and the trees on M.26 produce high quality fruit at a young age. Since it is prone to poor prop ability to soil, however, young trees require staking in windy locations. The rolC gene was introduced into M.26 by Agrobacterium tumefaciens LBA4404 harboring pBI121 to obtain its transformants with dwarfism and enhanced rooting ability. One regenerated transgenic line was confirmed by polymerase chain reaction (PCR) analysis and Southern blot analysis of genomic DNA for the existence of rolC gene. The characteristics of transgenic line in vitro were not significantly different from non-transgenic line except for the active root formation and lateral root number. The rolC transgenic line showed reduced stem length and increased root number in vitro. Rooting ability was examined in the isolated greenhouse after mound layering. Compared to non-transgenic M.26, rolC transgenic line showed significantly higher rooting ability. The transgenic line did not show any other observable variation in shoot phenotype compared with non-transgenic line excepting increased branching

Pulse wave velocity and ankle brachial index in adolescents with essential hypertension (본태성 고혈압 청소년에서 pulse wave velocity와 ankle brachial index에 대한 연구)

  • Joo, Sun Young;Cho, Ki Young;Cho, Su Jin;Hong, Young Mi
    • Clinical and Experimental Pediatrics
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    • v.49 no.7
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    • pp.769-776
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    • 2006
  • Purpose : Pulse wave velocity(PWV) and ankle brachial index(ABI) are not only noninvasive methods used to assess arterial stiffness in adults, but also useful, simple ways to estimate the severity of hypertension, end stage renal disease and atherosclerosis in adults. But there are few studies on PWV and ABI in adolescents and children. In this study, hypertensive adolescents were compared to normal individuals to find out the usefulness of PWV as the index of progress in the disease. Methods : 413 teenagers were divided into three groups. The first group consisted of adolescents that only had hypertension(Group 1 : 23 teenagers). The second group consisted of the adolescents who had hypertension with obesity(Group 2 : 17 teenagers), and the last group was the normal subjects(Group 3 : 328 teenagers). Weight, height and body mass index were measured. Simultaneous measurements of systolic, diastolic and average blood pressure were obtained from the four extremities. PWV, ABI, ejection time and preejection period were also measured. Results : Right brachial-ankle PWV was significantly higher in both group 1 and group 2 compared with group 3, and significantly higher in group 2 compared with group 1. Likewise, left brachial-ankle PWV also showed significantly higher values in both group 1 and group 2 compared with group 3, and also higher values in group 2 compared with group 1. Positive correlations were found between the systolic, diastolic, average blood pressure and PWV. There were also positive correlations between the blood pressure and weight, body mass index. Conclusion : Higher PWV was demonstrated in adolescents with essential hypertension compared to normal subjects. Follow-up study is needed to evaluate the progress.