• Journal of Plant Biotechnology
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• 제37권2호
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• pp.186-195
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• 2010

In most self-incompatible plant species, recognition of self-pollen is controlled by a single locus, termed the S-locus. The self-incompatibility (SI) system in Brassica is controlled sporophytically by multiple alleles at a single locus, designated as S, and involves cell-cell communication between male and female. Two highly polymorphic S locus genes, SLG (S locus glycoprotein) and SRK (S receptor kinase), have been identified, both of which are expressed predominantly in the stigmatic papillar cell. Gain-of-function experiments have demonstrated that SRK solely determines S haplotype-specificity of the stigma, while SLG enhances the recognition reaction of SI. The sequence analysis of the S locus genomic region of B. campestris (syn. rapa) has led to the identification of an anther-specific gene, designated as SP11/SCR, which is the male S determinant. Molecular analysis has demonstrated that the dominance relationships between S alleles in the stigma were determined by SRK itself, but not by the relative expression level. In contrast, the expression of SP11/SCR from the recessive S allele was specifically suppressed in the S heterozygote, suggesting that the dominance relationships in pollen were determined by the expression level of SP11/SCR. Furthermore, recent studies on recessive allele-specific DNA methylation of Brassica self-incompatibility alleles demonstrate that DNA methylation patterns in plants can vary temporally and spatially in each generation. In this review, we firstly present overview of self incompatibility system in Brassica and then describe dominance relationships in Brassica self- incompatibility regulated by allele-specific DNA methylation.

• 식물조직배양학회지
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• 제24권1호
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• pp.63-69
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• 1997

Erythropoietin(EPO)은 적혈구 모세포의 분화와 성장을 중재하는 당단백질이며 담배 식물체에서 재조합 사람 EPO를 생산하기 위해 CaMV 35S promoter를 갖는 발현 vector인 pBI$\Delta$GUS121, pBD$\Delta$ GUS121, pPEV-1을 이용하여 5.4kb의 EPO genomic DNA를 cloning 하였고 Agrobacterium tumefaciens에 의한 형질전환에 의해 Nicotiana tabacum (var. Xanthi)으로 도입되었다. Kanamycin을 포함하는 MS 배지에서 각각의 construct에 대하여 10 km 저항성 식물체들이 얻어졌다. 형질전환된 식물체의 게놈에 EPO genomic DNA의 정확한 결합은 polymerase chain reaction에 의해 332bP의 DNA 조각에 의해 확인되었으며 Northern blot 결과 1.8 kb의 전사체들이 식물체 잎에서 발현 축적되는 것이 확인되었다. Promoter의 수나 5'-UTS 서열에 의한 mRNA 양에는 변화가 없었지만 식물체 게놈에 결합된 위치 및 copy number에 의해 mRNA 수준에 영향을 주는 것으로 밝혀졌다. EPO 항체를 이용한 Western blot 결과 식물체에서 발현된 EPO 단백질의 크기는 동물세포에서 발현된 37kDa 보다 작은 30 kDa 이었다. 이는 식물체에서 modification(glycosylation) system은 동물세포에서와는 다르다는 것을 보여준다.

• 생약학회지
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• 제44권2호
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• pp.138-148
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• 2013

The three biochemical constituents pointed out as the bioactive principles of anticancer activity of Korean mistletoes, Viscum album var. colorantum, epiphyting to different host trees, different region, and different growth environments were compared. The characterizing compounds were lectin, which was glycoprotein reported has anticancer activity, homoflavoyadorinin-B, which was flavonoid glycoside reported as an antioxidant and oleanolic acid, as an anticancerous triterpenoid. We established the analysis methods for three characterizing compounds and determined by HPLC spectrophotometer, respectively. The average content of the lectin in V. album was 1.75 mg/g of dried plants and those of homoflavoyadorinin-B and oleanolic acid were 3.51 and 5.37 mg/g, respectively. The variations of contents of the three characterizing compounds according to epiphytic trees and growth region were observed but not found statistical significance clearly. We came to the conclusion that the contents of bioactive constituents of V. album except homoflavoyadorinin-B are not affected by epiphyting trees or growth environments.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.329-333
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• 2003

본 연구에서는 형질전환된 N. tabacum 배양에 있어서 배양 중반 저온으로의 변환이 세포에 미치는 영향과 hGM-CSF의 생산성 변화를 관찰하였다. 배양 중반 저온으로의 변환은 DCW의 증가와 세포크기의 감소를 보였다. Ascorbic acid의 첨가는 배양초기 세포 생존율의 감소를 완화시켰으며, 배양 중반 저온으로의 변환은 약간의 세포생존율 감소를 보였다. 저온으로의 변환, 저온 배양에서의 betaine 첨가, ascorbic acid 첨가 모두 배양 후반 세포 lysis 억제에 효과가 있었다. 배양 중반 저온으로의 변환시 배지내 단백질 분해 효소의 활성을 측정한 결과, 대조구 세포에 비해 낮은 단백질 분해 효소 활성을 나타내었다. 그로인해 배양 중반 이후 단백질 분해 효소에 의한 급격한 hGM-CSF 분해를 감소시킴으로써 상대적으로 대조구 세포에 비해 높은 hGM-CSF 생산성을 유지시켰다. Ascorbic acid를 첨가한 후 배양 도중 betaine(1 mM)을 첨가하여 저온으로 온도를 변환시, hGM-CSF의 생산성 대조구 세포에 비하여 최대 2.1배 까지 높게 유지시켰다.

• Journal of Photoscience
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• 제5권3호
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• pp.131-135
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• 1998

Using differential display reverse transcription technique, the present study attempted to isolate and characterize genes specifically expressed in cotyledons of Pharbitis nil Choisy cv. Violet during floral induction. A total of 107 bands specific to the inductive condition were initially obtained with 80 primer sets of 20 different arbitrary primers combined with 4 kinds of T12MN. In northern blot analysis with reamplified cDNAs as probes, three cDNAs were detected to be expressed specificcally in the induced cotyledon tissues, and designated PnFL-1, PnFL-2 and PnFL-3 , the size of which were 228 bp, 317 bp and 272 bp, respectively. A search for sequences similar to the decuced amino acid sequences was conducted using GenBank and EMBL database ; seequence encoded by PnFL-1 had 29% identity with the clone of Arabidopsis thaliana similiar to reverse trascriptase (Genbank Acc. N0.3047086), PnFL-2 shared 50% identity with hydroxiyproline-rich glycoprotein of Glycine max(GenBank Acc. No.347455), and PnFL-3 had 46% identity with TAMU 4. Thaliana genomic clone T23E16 (GenBank Acc. No.B67574). None of them was known gene in the plant system up to date, implying that the fragments may comprise parts of genes which are associated with the floral induction in Pharbitis nil.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.133.1-133
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• 2003

Gummy stem blight, caused by Didymella bryoniae occurs exclusively on cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. Through this study, we optimized cultural conditions for mass-production of pycnidiospore by Metal Halide Lamp irradiation. In brief, the mycelial was cultured at $26^{\circ}C$ on PDA, for 2 days under the darkness, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$, a great number of pycnidia was simultaneously formed. Thus produced pycnidiospores were used as immunogen. From fusions of myeloma cell (v-653) with splenocytes from immunifed mice were car ried out. And, two hybridoma cell lines that recognized the immunogen Didymella bryoniae were obtained. One Monoclonal Antibody, Db1, recognized the supernatant and the other monoclonal antibody, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid two MAb Db1 and Db15, were immunotyped and identified as IgG1 and IgG2b, respectively. Titer of MAb Db1 and MAb Db15 was measured absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with Didymella bryoniae but none reacted with other of fungi and CMV, CGMMV Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{3}$ well by indirect ELISA characterization of the MAb Db1, Db15 antigen by heat and protease treatments show that the epitope recognized by the MAb Bb1, Db15 were a glycoprotein.

• The Plant Pathology Journal
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• 제19권5호
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• pp.260-265
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• 2003

Gummy stem blight caused by Didymella bryoniae occurs exclusively in cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. In this study, cultural conditions for the mass-production of pycnidiospore by Metal Halide (MH) lamp irradiation were maximized. The mycelia were cultured at $26^{\circ}C$ on PDA for 2 days under dark condition, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$. Results show that a great number of pycnidia were simultaneously formed. The pycnidiospores produced were then used as immunogen. Fusions of myeloma cell (v-653) with splenocytes from immunized mice were carried out. Two hybridoma cell lines that recognized the immunogen D. bryoniae were obtained. One monoclonal antibody (MAb), Dbl, recognized the supernatant while another MAb, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid of the two MAb, Dbl and Db15, the immunotypes of which were identified as IgG1 and IgG2b, respectively. Titers of MAb Dbl and MAb Db15 were measured and the absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with D. bryoniae but none reacted with other viral isolates, Cucumber mosaic virus and Cucumber green mottle mosaic virus. Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{-3}$/well by indirect ELISA. Characterization of the MAbs Dbl, Db15 antigen by heat and protease treatments, which suggests that the epitope recognized by these two MAbs was glycoprotein.

• Journal of Plant Biotechnology
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• 제34권2호
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• pp.145-152
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• 2007

Lactoferrin is an 80-kDa iron-binding glycoprotein known to exert many biological activities, such as facilitating iron absorption and having antimicrobial and anti-inflammatory effects. Rice can be a useful target for edible food plants to introduce human lactoferrin, because it has lower allergenicity and is likely to be safer than microorganisms or transgenic animals. A cDNA fragment encoding human lactoferrin (HLF) driven by the maize polyubiquitin promoter, along with herbicide resistance gene (bar) driven by CaMV 35S promoter, was introduced into rice (Oryza sativa L. cv. Dong Jin) using the Agrobacterium -mediated transformation system. Putative transformants were initially selected on the medium containing bialaphos. The stable integration of the bar and HLF genes into transgenic rice plants was further confirmed through polymerase chain reaction (PCR) and Southern blot analyses. The expression of the full length HLF protein from various tissues such as grains and young leaves of transgenic rice was verified by Western blot analysis. Analysis of progeny also demonstrated that introduced genes were stably inherited to the next generation at the Mendelian fashion.

• Genomics & Informatics
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• 제14권3호
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• pp.96-103
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• 2016

The influenza A (H1N1) virus, also known as swine flu is a leading cause of morbidity and mortality since 2009. There is a need to explore novel anti-viral drugs for overcoming the epidemics. Traditionally, different plant extracts of garlic, ginger, kalmegh, ajwain, green tea, turmeric, menthe, tulsi, etc. have been used as hopeful source of prevention and treatment of human influenza. The H1N1 virus contains an important glycoprotein, known as neuraminidase (NA) that is mainly responsible for initiation of viral infection and is essential for the life cycle of H1N1. It is responsible for sialic acid cleavage from glycans of the infected cell. We employed amino acid sequence of H1N1 NA to predict the tertiary structure using Phyre2 server and validated using ProCheck, ProSA, ProQ, and ERRAT server. Further, the modelled structure was docked with thirteen natural compounds of plant origin using AutoDock4.2. Most of the natural compounds showed effective inhibitory activity against H1N1 NA in binding condition. This study also highlights interaction of these natural inhibitors with amino residues of NA protein. Furthermore, among 13 natural compounds, theaflavin, found in green tea, was observed to inhibit H1N1 NA proteins strongly supported by lowest docking energy. Hence, it may be of interest to consider theaflavin for further in vitro and in vivo evaluation.

• Journal of Plant Biotechnology
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• 제32권3호
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• pp.209-215
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• 2005

락토페린은 철 결합 당단백질로서 항 미생물활성과 면역강화와 같은 생리활성 기능을 가지고 있다. 본 연구는 배양 세포 고 발현 SWPA2 promoter를 이용하여 인체락토페린(hLf)을 생산하는 형질전환 가시오갈피 배양세포주 개발에 관한 것이다. 형질전환에 이용된 벡터는 산화스트레스 유도성 SWPA2 promoter의 조절 하에서 hLf이 소포체로 targeting되도록 제작된 SWPA2pro::ER-hLf/pCAMBIA이다. hLf을 생산하는 각 형질전환 배양세포들은 PCR과 Southern분석을 통해 hLf 유전자가 가시오갈피 게놈내로 성공적으로 도입되었음을 확인하였으며, western blot과 ELISA를 통해 형질전환 가시오갈피 배양 세포주에서 hLf 단백질이 활성이 있음을 확인하였다. 형질전환 가시오갈피 배양세포에서 hLf 단백질의 함량은 세포배양이 진행될수록 증가하여 정지기 때 가장 높았으며 전체 수용성 단백질의 약 3%를 차지하였다. 따라서 본 연구에서 개발된 인체락토페린을 고생산하는 약용식물 가시오갈피 배양 세포주는 산업적으로 이용될 수 있을 것으로 기대된다.