• 생명과학회지
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• 제7권4호
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• pp.270-275
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• 1997

In order to produce betacyanins from sugar beet(Beta vulagris L.) in vitro, callus induction, shoot formation, root formation, betacyanin contents and protein contents determined from callus which was induced cotyledons of sugar beet seedling under an addition of NAA and BAP on the MS medium. The results were summarized as follows; The combination 3.0mg/l NAA and 1.0mg/l BAP treatment showed the most high callus induction rate, betacyanin and protein contents. The combination NAA and BAP treatments were not shoot formation, but BAP treatments showed high root formation rate. But high concentrations of BAP have not shown root formation.

• 생명과학회지
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• 제11권3호
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• pp.254-258
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• 2001

The biochemical change during regeneration of perilla callus were investigated by comparing total protein and lipid contents, protein band pattern in SDS-PAGE, and fatty acid composition in the calli cultured for various period(0, 1, 3, 5 and 6 weeks) Calli were induced from cotyledon and hypocotyl explants of peplants of perilla on perilla on MS medium containing BA(0.5 mg/L) and NAA(0.5mg/L). The protein contents reached the peak at 3 weeks after induction of calli, and then was decreased. Total lipid contents was decreased as the culture period increased. The band pattern of polypeptides showed that 30KD and 45KD polypeptides and 22KD and 45KD polypetides were major proteins in the cotyledon and hypocotyl explants, respectively. However increase of culture period only 30KD protein was highly accumulated.

• 생명과학회지
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• 제9권1호
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• pp.29-34
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• 1999

들깨로부터 callus의 유기 및 재분화를 위해 MS 기본배지에 생장조정제 NAA(0.1, 0.5, 1.0, 및 2.0mg/$\ell$)와 BA(0.5, 1.0 및 1.5mg/$\ell$)를 혼용하여 자엽과 하배축 조직을 치상하고 3, 4, 5 및 6주로 나누어 재분화과정과 구성성분의 변화를 조사한 결과, BA 0.5mg/$\ell$과 NAA 0.5mg/$\ell$을 첨가한 배지에서 자엽과 하배축 모두 callus의 유기 및 재분화 정도가 좋았고, 자엽절편의 뿌리 발생은 BA 0.5mg/$\ell$과 NAA 0.1mg/$\ell$을 첨가한 배지에서만 발생하였다. 배양기간의 경과에 따라 단백질함량은 증가한 반면 RNA 함량은 감소하였고, polypeptide pattern은 배양전 자엽조직에서는 30KD과 45KD의 polypeptide가 진하게 나타났고, 30KD polypeptide는 배양기간의 경과에 따라 증가 하였다.

• BMB Reports
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• 제30권1호
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• pp.21-25
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• 1997

The growth. freezing resistance and electrophoretic protein patterns of carrot callus cultures were investigated following treatment with NaCl for various' intervals at 20$^{\circ}C$. Following 7 day exposure to 250 mM NaCl. freezing tolerance increased, which was measured by 2.3.5-triphenyl tetrazolium chloride (TTC) assay and fresh weight was reduced compared to control cells. Changes of electrophoretic patterns of total and boiling stable proteins were investigated using one or two dimensional gel system. Several proteins with molecular weight of 43 and 21 kDa increased by NaCl treatment. The most prominent change was detected in 21 kDa protein. The steady state level of this protein increased in NaCl treated cells, but decreased in control cells. Twenty one kDa protein was detected only in the NaCl treated cell when boiling stable protein was analyzed. The isoelectric point of 21 kDa protein was identified as 5.7. The timing of increase of 21 kDa protein was correlated to freezing resistance which implied the role of this protein in the induction of freezing resistance of the cell.

• 한국육종학회지
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• 제49권3호
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• pp.170-179
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• 2017

분자농업은 식물을 일종의 공장개념으로 확대 적용하여 산업적으로 가치가 높은 유용물질들을 대량생산하는 분야로 최근 많은 각광을 받고 있다. 그 중에서 벼 캘러스를 이용한 단백질 발현 시스템은 대량 배양이 가능하고 목적 단백질의 높은 발현율로 인한 산업화가 가능한 기술이다. 본 연구는 이러한 벼(Oryza sativa L.) 캘러스의 활용도를 높이기 위한 효율적인 형질전환 시스템을 구축하기 위해 수행되었다. 형질전환에 이용된 종자의 종피 제거 시 손을 이용함으로써 캘러스 유도율을 높여주었고, 6년 정도의 오래된 종자에서도 원활한 캘러스 유도가 가능하였다. 목적 유전자가 도입된 캘러스의 선발은 최소한 3주 이상의 배양기간을 필요로 하였고 가장 효율적인 것은 한번의 계대를 포함한 6주 배양 후 선발하는 것이었다. 이러한 선발은 유전자가 식물세포의 genomic DNA 안에 안정적으로 삽입되어 이루어졌음을 서던 블롯 분석 및 후대검정 등을 통하여 확인할 수 있었다. 이러한 장기적이고 대량의 배양이 가능한 벼 캘러스의 효율적인 선발 시스템은 벼를 이용한 유용 재조합 단백질의 산업화에 실속있는 기술로 기여할 수 있을 것이다.

• Journal of Plant Biotechnology
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• 제35권2호
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• pp.133-140
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• 2008

DNA와 histone 단백질의 변형은 식물 발달에 상당히 중요한 역할을 하는 것으로 알려져 있다. 식물 조직 배양 및 식물 발달 단계에서 methylation의 영향을 알아보고자 벼 종자로부터 캘러스 형성 및 식물체 재분화 단계에서 demethylation 물질인 5-azacytidine을 처리하여 유전자 발현 양상을 분석하였다. 식물체로의 재분화 능력이 있는 벼 배상체 캘러스는 5-azaC가 첨가된 H6A 배지에서는 형성되지 않았으며 갈색을 띠는 캘러스가 형성되었다. 또한 정상적인 캘러스를 5-azaC가 첨가된 MSRA 재분화 배지에서 배양했을 때도 대조구와는 달리 식물체 재분화는 이루어지지 않았다. 이러한 결과는 5-azaC가 정상적인 배상체 캘러스 및 shoot 분화에 부정적인 영향을 미친다는 것을 나타냈으며 따라서 DNA methylation이 식물 조직배양에서의 정상적인 세포 dedifferentiation과 differentiation에 필수 요인이라는 것을 알 수 있었다. 벼 캘러스 형성 및 재분화 과정 동안의 methylation 영향을 알아보고자 각 단계별로 5-azaC를 처리 후 $GeneFishig^{TM}$ DEG와 DNA chip을 사용하여 유전자 발현 양상을 분석하였다. Epigenetic regulation, 전자전달, 핵산대사, 스트레스 반응에 관여하는 일부 유전자들의 발현이 증가하거나 감소하는 것을 알 수 있었다. 발현 차이가 있는 일부 유전자를 클로닝하여 확인하였고 RT-PCR 및 northern 분석으로 각 단계에서의 발현 차이를 할인하였다.

• Journal of Plant Biotechnology
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• 제30권4호
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• pp.315-321
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• 2003

고비사막 접경지역의 식물인 Artermisia adamsii의 내건성 및 내동성 증진을 위해 조직배양과 CLP및 Dhn5유전자의 형질전환을 수행하였다. 다양한 호르몬 농도 조건에서의 성장을 확인한 결과, 0.05mg/L의 NAA와 0.5mg/L의 BAP조건과 0.1mg/L의 NAA와 0.5mg/L의 BAP가 포함된 배지의 암조건 하에서 최적의 캘러스 생장을 확인하였으며 유전자 도입 및 유전자의 발현이 확인된 캘러스가 세포 수준에서도 내건성 및 내동성이 증진되었음을 확인하였다. 그러나 Atremisia속 다른 식물과 다르게 조직절편에서 직접 기관분화를 유도하거나 캘러스를 통한 식물체 재생에 어려움이 있어 식물체 수준에서 형질전환에 따른 환경스트레스 내성의 증진을 확인하지 못하고 있다. 앞으로 진행될 A. adamsii의 식물체 재생에 대한 연구를 통해 내동성 및 내건성이 증진된 식물체를 육성하여 고비사막 지역적응력을 조사할 예정이다.

• The Plant Pathology Journal
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• 제15권1호
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• pp.38-43
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• 1999

Mutants of the monopartite geminivirus beet curly top virus (BCTV) have been screened for infectivity, systemic movement, replication and symptom development in Arabidopsis thaliana. As known by coding for coat protein, R1 mutant was not infectious and did not move systemically. R2, R3 and L2/L3 mutants produced milder symptoms compared to wild type BCTV but the infectivity was reduced by 40% to 60%. R2 ORF is thought to be involved in the regulation of ssDNA and dsDNA accumulation because only dsDNA was accumulated on R2-infected organs. Disruption of ORF L4 resulted in reduced infections, but the viral DNA was accumulated in infected organs from roots to shoot tips as much as wild type BCTV on Sei-O. In addition, 4 mutants did not produce callus-like tissues on infected organs, suggesting that L4 ORF may play a role in the induction of host cell divisions by virus infection. This result was supported by the patterns of mRNA expression and promoter analysis of the cell cycle marker gene, cycl, on Arabidopsis. cycl mRNA was accumulated on symptomatic organs by wild type BCTV infections but not by L4 mutant. We conclude that the BCTV L4 ORF is essential for symptom developments, specially callus-like formation on infected organs.

• KSBB Journal
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• 제6권3호
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• pp.289-297
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• 1991

The insecticidal protein gene in the pKL-20-1 clone derived from Bacillus thuringiensis serovar. kurstaki plasmid was subcloned in the plant shuttle vector, pGA643. The 7.3 kb fragment was cloned in the BglII and Hpal sites of pGA643 vector and expressed in E. coli S17-1, which produced insecticidal proteins killing Bombyx mori larvae. The clone was named pHL-20. The protoplast formation, calli induction and plantlet regeneration of Nicotiana tabacum was carried out. A tremendous number of mesophyll protoplasts of N. tabacum were formed, up to 7$\times$105 protoplast per ml, for 20 hours in darkness in the enzyme solution of 0.5% cellulase and 0.1% macerosin, pH 5.8. The viabilities of the protoplasts were maintained above 80% for 6 days in the media containing 2mg/1 of NAA and 1mg/1 of kinetin. Calli were induced from the protoplasts and leaves of the N. tabacum on MS medium containing 0.5mg/1 BAP. Under the culture conditions the protoplasts underwent repeated cell division into calli. Plantlets were regenerated from callus cultures derived from protoplast and leaves. Shoots were induced in a medium containing 1mg/1 of BAP.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권2호
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• pp.154-159
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• 2006

Interleukin-18 (IL-18), otherwise known as interferon-gamma-inducing factor (IGIF), is one of several well characterized and important cytokines that contribute to host defenses. The complementary DNA (cDNA) of mature human interleukin-18 gene (hIL-18) was fused with the signal peptide of the rice amylase 1A gene (Ramy1A) and introduced into the plant expression vector under the control of a duplicated CaMV 35S promoter. The recombinant plasmid was transformed into tobacco (Nicotiana tabacum L. cv Havana) using the Agrobacterium-mediated transformation method. The integration of the hlL-18 gene into the genome of transgenic tobacco plants was confirmed by polymerase chain reaction (PCR) amplification and its expression was observed in the suspension cells that were derived from the transgenic plant callus by using Northern blot analysis. The hlL-18 protein was detected in the extracts of the transgenic callus and in the medium of the transgenic tobacco suspension culture by using immunoblot analysis. Based upon enzyme-linked immunosorbant assay (ELISA) results, the expression level of the hlL-18 protein approximated $166{\mu}g/L$ in the suspension culture medium. Bioassay results from the induction of $interferon-{\gamma}$ from a KG-1 cell line indicated that the hlL-18 secreted into the suspension culture medium was bioactive.