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Research status of the development of genetically modified papaya (Carica papaya L.) and its biosafety assessment (GM 파파야 개발 및 생물안전성 평가 연구 동향)

  • Kim, Ho Bang;Lee, Yi;Kim, Chang-Gi
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.171-182
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    • 2018
  • Papaya (Carica papaya L.) is one of the crops widely planted in tropical and subtropical areas. The papaya fruit has low calories and are plentiful in vitamins A and C and in minerals. A major problem in papaya production is a plant disease caused by the papaya ringspot virus (PRSV). The first PRSV-resistant GM papaya expressing a PRSV coat protein gene was developed by USA scientists in 1992. The first commercial GM papaya cultivars derived from the event was approved by the US government in 1997. Development of transgenic papayas has been focused on vaccine production and limited agricultural traits, including insect and pathogen resistance, long shelf life, and aluminum and herbicide tolerance. Approximately 17 countries, including the USA and China, produced transgenic papayas and/or commercialized them, which provoked studies on biosafety assessment and development of GM-detection technologies. For the biosafety assessment of potential effects on human health, effects of long-term feeding to model animals have been studied in terms of toxicity and allergenicity. Studies on environmental safety assessment include influence on soil-microbial biodiversity and transfer to soil bacteria of GM selection markers. Many countries, such as Korea, the European Union, and Japan, that have strict regulations for GM crops have serious concerns about unintended introduction of GM cultivars and food commodities using unauthorized GM crops. Transgene- and/or GM event-specific molecular markers and technologies for genomics-based detection of unauthorized GM papaya have been developed and have resulted in the robust detection of GM papayas.

Isolation and functional analysis of three microsomal delta-12 fatty acid desaturase genes from Camelina sativa (L.) cv. CAME (카멜리나 (Camelina sativa L. cv. CAME)로부터 3 microsomal delta-12 fatty acid desaturase 유전자들의 분리 및 기능 분석)

  • Kim, Hyojin;Go, Young Sam;Kim, Augustine Yonghwi;Lee, Sanghyeob;Kim, Kyung-Nam;Lee, Geung-Joo;Kim, Gi-Jun;Suh, Mi Chung
    • Journal of Plant Biotechnology
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    • v.41 no.3
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    • pp.146-158
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    • 2014
  • Camelina sativa that belongs to Brassicaceae family is an emerging oilseed crop. Camelina seeds contain approximately 40% storage oils per seed dry weight, which are useful for human and animal diets and industrial applications. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. In this study, three CsFAD2 genes (CsFAD2-1, CsFAD2-2 and CsFAD2-3.1) were isolated from developing seeds of Camelina sativa (L.) cv. CAME. The nucleotide and deduced amino acid sequences of three CsFAD2 genes were compared with those from dicotyledon and monocotyledon plants including Camelina cultivars Sunesone and SRS933. Three histidine motifs (HECGH, HRRHH, and HVAHH) required for FAD activity and a hydrophobic valine or isoleucine residue, which is a SNP (single nucleotide polymorphism) marker related with enzyme activity are well conserved in three CsFAD2s. The expressions of CsFAD2-1 and CsFAD2-3.1 were ubiquitously detected in various Camelina organs, whereas the CsFAD2-2 transcripts were predominantly detected in flowers and developing seeds. The contents of oleic acids decreased, whereas the amounts of linoleic acid increased in dry seeds of transgenic fad2-2 lines expressing each CsFAD2 gene compared with fad2-2 mutant, indicating that three CsFAD2 genes are functionally active. The isolated CsFAD2 genes might be applicable in metabolic engineering of storage oils with high oleic acids in oilseed crops.

Effects of Garcinia cambogia Extract on the Adipogenic Differentiation and Lipotoxicity (가르시니아 캄보지아 추출물의 지방세포 분화 및 지방 독성에 미치는 영향)

  • Kang, Eun Sil;Ham, Sun Ah;Hwang, Jung Seok;Lee, Chang-Kwon;Seo, Han Geuk
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.411-416
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    • 2013
  • This study aimed to examine the mechanisms underlying the effects of Garcinia cambogia extract on the adipogenic differentiation of 3T3-L1 cells and long-chain saturated fatty acid-induced lipotoxicity of HepG2 cells. 3T3-L1 preadipocytes, mouse embryonic fibroblast-adipose like cell line, were treated with MDI solution (0.5 mM IBMX, 1 ${\mu}M$ dexamethasone, 10 ${\mu}g/mL$ insulin) to generate a cellular model of adipocyte differentiation. Using this cellular model, the anti-obesity effect of Garcinia cambogia extract was evaluated. MDI-induced lipid accumulation and expression of adipogenesis-related genes were detected by Oil red O staining, Nile Red staining, and Western blot analysis. Effects Garcinia cambogia extract on palmitate-induced lipotoxicity was also analyzed by MTT assay, LDH release, and DAPI staining in HepG2 cells. Garcinia cambogia extract significantly suppressed the adipogenic differentiation of preadipocytes and intracellular lipid accumulation in the differentiating adipocytes. Garcinia cambogia extract also markedly inhibited the expression of peroxisome proliferator- activated receptor ${\gamma}2$ ($PPAR{\gamma}2$), CCAT/enhancer-binding protein ${\alpha}$ ($C/EBP{\alpha}$), and adipocyte protein aP2 (aP2). In addition, Garcinia cambogia extract significantly attenuated palmitate-induced lipotoxicity in HepG2 cells. Palmitateinduced cellular damage and reactive aldehydes were also significantly reduced in the presence of Garcinia cambogia extract. These findings suggest that the Garcinia cambogia extract inhibits the adipogenic differentiation of 3T3-L1 preadipocytes, probably by regulating the expression of multiple genes associated with adipogenesis such as $PPAR{\gamma}2$, $C/EBP{\alpha}$, aP2, and thereby modulating fatty acid-induced lipotoxicity to reduce cellular injury in hepatocytes.

Effect of submerged culture of Ceriporia lacerata mycelium on GLUT4 protein in db/db mouse (db/db 마우스에서 Ceriporia lacerata 균사체 배양액이 GLUT4 발현에 미치는 영향)

  • Shin, Eun Ji;Kim, Ji-Eun;Kim, Ji-Hye;Park, Yong Man;Yoon, Sung Kyoon;Jang, Byeong-Churl;Lee, Sam-Pin;Kim, Byoung-Cheon
    • Food Science and Preservation
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    • v.22 no.6
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    • pp.893-900
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    • 2015
  • In this study, we evaluated the antidiabetic effect of a submerged culture of Ceriporia lacerata mycelium (CL01) on hematological indices, as well as protein and mRNA expression of the insulin-signaling pathway, in db/db mice. After CL01 was administrated for 4 weeks, blood glucose levels decreased consistently, and plasma insulin and c-peptide levels each decreased by roughly 55.8%, 40% of those in the negative control (p<0.05). With regard to HOMA-IR, an insulin resistance index, insulin resistance of the CL01-fed group improved over that of the negative control group by about 62% (p<0.05). In addition, we demonstrated that the protein expression levels of pIR, pAkt, pAMPK, and GLUT4 and the mRNA expression levels of Akt2, IRS1, and GLUT4 in the muscle cells of db/db mice increased in the CL01-fed group compared to the corresponding levels in the control group. These results demonstrate that CL01 affects glucose metabolism, upregulates protein and gene expression in the insulin-signaling pathway, and decreases blood glucose levels effectively by improving insulin sensitivity. More than 90% of those who suffer from type 2 diabetes are more likely to suffer from hyperinsulinemia, hypertension, obesity, and other comorbidities because of insulin resistance. Therefore, it is possible that CL01 intake could be used as a fundamental treatment for type 2 diabetes by lowering insulin resistance, and these results may prove be useful as basic evidence for further research into the mechanisms of a cure for type 2 diabetes.

Characterization of an Extracellular Xylanase from Bacillus sp. HY-20, a Bacterium in the Gut of Apis mellifera (꿀벌(Apis mellifera)의 장내 세균인 Bacillus sp. HY-20이 분비하는 Xylanase의 특성)

  • Lee, Lan-Hee;Kim, Do-Young;Han, Mi-Kyoung;Oh, Hyun-Woo;Ham, Su-Jin;Park, Doo-Sang;Bae, Kyung-Sook;Sok, Dai-Eun;Shin, Dong-Ha;Son, Kwang-Hee;Park, Ho-Yong
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.332-338
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    • 2009
  • A xylan-decomposing bacterium, HY-20, was isolated from the gut of a honeybee, Apis mellifera, and identified as Bacillus sp. The extracellular GH11 xylanase (XylP) gene (687-bp) of strain HY-20 encoded a protein of 228 amino acids with a deduced molecular mass of 25,522 Da and a calculated pI of 9.33. The primary structure of XylP was 97% identical to that of B. pumilus xylanase (GenBank accession no.: AY526092) that has not been characterized yet. The recombinant His-tagged enzyme (rXylP) overexpressed in Escherichia coli BL21 harboring pET-28a(+)/xylP was purified to electrophoretic homogeneity by cation exchange and gel permeation chromatographies. The purified enzyme exhibited the highest catalytic activity toward birchwood xylan at pH 6.5 and $50^{\circ}C$ and retained approximately 50% of its original activity when pre-incubated at $55^{\circ}C$ for 15 min. The recombinant enzyme was completely inactivated by $Hg^{2+}$ (1 mM) and N-bromosuccinimide (5 mM), while its activity was slightly stimulated by approximately 10% in the presence of $Mn^{2+}$ (1 mM), $Fe^{2+}$ (1 mM), and sodium azide (5 mM). rXylP was able to efficiently degrade various polymeric xylose-based substrates but PNP-sugar derivatives and glucose-based polymers were not susceptible to the enzyme.

Antiviral Properties of Probiotic Mixtures against Rotavirus in the Rat (랫드에서 로타바이러스에 대한 유산균혼합물의 항 바이러스활성)

  • Park, Jae Eun;Lee, Do Kyung;Kim, Min Ji;Kim, Kyung Tae;Choi, Kyung Soon;Seo, Jae Goo;Ha, Nam Joo
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.296-301
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    • 2014
  • Rotavirus is a major cause of acute gastroenteritis in young children in developed and developing countries. The use of probiotics for the treatment of gastrointestinal diseases is both safe and easily accessible. In this study, we evaluated the anti-rotaviral activities of probiotic mixtures in a Sprague-Dawley rat. 24 litters with their dams were randomly assigned to four groups; placebo, phosphate buffered saline (PBS), and two probiotic mixture (PRO-1 and PRO-2) groups. All rats were inoculated with rotavirus at dose of 8 log plaque forming units per rat at 5 days old. Animals in the PRO-1 and PRO-2 groups were orally administered probiotic mixtures 1 or 2, respectively, at a dose of 8 log colony forming units daily during 4 days. For control purposes, placebo and PBS groups were orally administered the same amount of placebo (containing maltose and polydextrose) or PBS once daily for 4 days, respectively. Antiviral analysis was performed by real-time quantitative PCR (RT-qPCR) and observing intestinal villi. As a result, weights of small intestines were greater in the PRO-1, PRO-2 groups than in control groups. Villi were short and villous epithelial necrosis was exhibited in control groups, but these morphological changes were not observed in PRO-1, PRO-2 treated rats. RT-qPCR analysis showed that VP7 gene level of rotavirus in fecal samples and small intestinal epithelial cells were lower in the PRO-1 and PRO-2 groups. These findings suggest that probiotic mixtures may be useful probiotics for the treatment of or as alternative therapies for rotaviral gastroenteritis.

Expression of Immortalization-upregulated Proteins-2 (IMUP-2) in Placenta (태반 내 Immortalization-upregulated Proteins-2 (IMUP-2) 발현)

  • Jeon, Su-Yeon;Lee, Hyun-Jung;Jung, Hyun-Min;Kim, Jin-Kyeoung;Kim, Gi-Jin
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.3
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    • pp.163-174
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    • 2009
  • Objectives: Members of the immortalization-upregulated protein (IMUP) family are nuclear proteins implicated in SV40-mediated immortalization and cellular proliferation, but the mechanisms by which their expression is regulated are still unknown in placenta. To investigate to expression and functions of IMUPs in placenta, we conducted to compare IMUPs expression in normal and preeclamptic placenta tissues and analyzed the function of IMUP-2 in HTR-8/SVneo trophoblast cells after IMUP-2 gene transfection. Methods: The expression of IMUPs was analyzed in placental tissues from the following groups of patients (none underwent labor): 1) term normal placenta (n=15); 2) term with preeclamptic placeneta (n=15); and 3) pre-term with preeclamptic placenta (n=11) using semi-quantitative RT-PCR, RNA in situ hybiridization, immunohistochemistry, and Western blot. In order to evaluate the function of IMUP-2 in HTR-8/SVneo trophoblast cells, IMUP-2 plasmids were transfected into HTR-8/SVneo trophoblast cells for 24 hours. Results: We observed that IMUPs are mainly expressed in the syncytiotrophoblasts and syncytial knot of placental villi. The expression of IMUP-1 was not differences between normal and preeclamptic placenta tissues. However, IMUP-2 expression was significantly higher in preterm preeclamptic placenta tissues than in normal placenta tissues without labor (p<0.001). Furthermore, we confirmed overexpression of IMUP-2 induced apoptosis in HTR-8/SVneo trophoblast cells through up-regulation of pro-apoptotic proteins. Conclusions: These results suggest that the expression of IMUP-2 is involved in placental development as well as increased IMUP-2 expression is associated with preeclampsia through the inducing of trophoblast apoptosis.

Correlation and Combining Ability related to the Growth Characteristics in F1 Hybrids by Diallel Cross of Viola tricolor (팬지의 일대 잡종에서 생육관련 형질의 상관관계 및 조합능력)

  • Song, C.Y.;Hong, K.H.;Kang, Y.K.;Lee, K.H.;Kim, J.C.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.4 no.1
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    • pp.70-79
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    • 2002
  • This study was conducted to determine correlations and combining abilities of leaf lengths, leaf widths, petiole lengths, number of branches, number of leaves, leaf areas, fresh weights, dry weights and number of flowers in F1 crosses made with the partial seven-parent diallel cross in Viola tricolor. Leaf lengths and leaf widths showed highly positive correlation with petiole length and negative correlation with number of branches, number of leaves and number of flowers. There was positive correlation between the number of leaves and flowers as well as between leaf area and fresh weight. Mean squares of general combining ability(GCA) and specific combining ability(SCA) were highly significant for all the parameters. Variance component values of SCA were greater than those of GCA for all the parameters except leaf length, implying preponderance of non-additive gene actions for these characters. The lines C and G for leaf lengths and widths, the lines A and F for number of leaves, the lines A, B and G for leaf areas, and the lines B and F for number of flowers showed relatively high GCA effects. The crosses of A×B and B×D exhibited high SCA effects on increasing leaf lengths, leaf widths, number of leaves, fresh weights and number of flowers. And also the crosses of B×E and D×G exhibited high SCA effects on decreasing leaf lengths, leaf areas and fresh weights as well as increasing number of flowers. The broad sense heritabilities of most characters were high compared with the narrow sense one. Those of leaf length, leaf width, petiole length and number of leaves were high in both the broad and narrow sense heritabilities.

Ethanol extract of Aster glehni exhibits anti-inflammatory and anti-oxidant effects in RAW 264.7 cells and Caenorhabditis elegans (섬쑥부쟁이 에탄올 추출물이 대식세포와 예쁜꼬마선충에서의 항염증 및 항산화 효과 )

  • Mi-Kyung Seo;Han-Na Chu;Da-Bin Lee;Haeng-Ran Kim;In-Seon Hwang;Yong-Jin Jeong;Sung-Ran Yoon;Seok-Seong Kang;Kyeong-A Jang;Min-Sook Kang
    • Food Science and Preservation
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    • v.30 no.6
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    • pp.1095-1106
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    • 2023
  • This study investigated the anti-oxidative and anti-inflammatory effects of Aster glehni (AG) extract in RAW 264.7 cells and Caenorhabditis elegans. The total polyphenol and flavonoid contents were higher in the ethanol extracts than in the hot water extracts. As a result of measuring the moisture contents (%) and extraction yields (%) of AG and drying A. glehni for processing (DAG), 70% ethanol, which has the highest percentage of extraction yield, was selected as the final solvent. DPPH radical scavenging activity showed higher antioxidant activity of ethanol extracts of DAG than AG. The cytotoxicity assay of the AG or DAG ethanol extracts was treated at different concentrations (25, 50, and 100 ㎍/mL), and cell viability rates were higher than 80% at all concentrations. The LPS-stimulated nitric oxide (NO) production in RAW 264.7 was significantly reduced at all concentrations of AG and DAG groups. As a result of measuring the gene expression of iNOS, which induces NO production, the AG or DAG group decreased by 33% and 32%, compared with the phosphate buffer saline (PBS) group. Under inflammatory stress conditions, the survival rate of C. elegans treated with AG or DAG ethanol extract with LPS showed concentration-dependent improvement in survival rate compared with the PBS group. Considering these results, AG could potentially be developed as an antioxidant and anti-inflammatory functional food material.