• Title/Summary/Keyword: Gene Technology

Search Result 3,640, Processing Time 0.04 seconds

Metabolic Engineering for Resveratrol Derivative Biosynthesis in Escherichia coli

  • Jeong, Yu Jeong;Woo, Su Gyeong;An, Chul Han;Jeong, Hyung Jae;Hong, Young-Soo;Kim, Young-Min;Ryu, Young Bae;Rho, Mun-Chual;Lee, Woo Song;Kim, Cha Young
    • Molecules and Cells
    • /
    • v.38 no.4
    • /
    • pp.318-326
    • /
    • 2015
  • We previously reported that the SbROMT3syn recombinant protein catalyzes the production of the methylated resveratrol derivatives pinostilbene and pterostilbene by methylating substrate resveratrol in recombinant E. coli. To further study the production of stilbene compounds in E. coli by the expression of enzymes involved in stilbene biosynthesis, we isolated three stilbene synthase (STS) genes from rhubarb, peanut, and grape as well as two resveratrol O-methyltransferase (ROMT) genes from grape and sorghum. The ability of RpSTS to produce resveratrol in recombinant E. coli was compared with other AhSTS and VrSTS genes. Out of three STS, only AhSTS was able to produce resveratrol from p-coumaric acid. Thus, to improve the solubility of RpSTS, VrROMT, and SbROMT3 in E. coli, we synthesized the RpSTS, VrROMT and SbROMT3 genes following codon-optimization and expressed one or both genes together with the cinnamate/4-coumarate:coenzyme A ligase (CCL) gene from Streptomyces coelicolor. Our HPLC and LC-MS analyses showed that recombinant E. coli expressing both ScCCL and RpSTSsyn led to the production of resveratrol when p-coumaric acid was used as the precursor. In addition, incorporation of SbROMT3syn in recombinant E. coli cells produced resveratrol and its mono-methylated derivative, pinostilbene, as the major products from p-coumaric acid. However, very small amounts of pterostilbene were only detectable in the recombinant E. coli cells expressing the ScCCL, RpSTSsyn and SbROMT3syn genes. These results suggest that RpSTSsyn exhibits an enhanced enzyme activity to produce resveratrol and SbROMT3syn catalyzes the methylation of resveratrol to produce pinostilbene in E. coli cells.

Kinetics of a Cloned Special Ginsenosidase Hydrolyzing 3-O-Glucoside of Multi-Protopanaxadiol-Type Ginsenosides, Named Ginsenosidase Type III

  • Jin, Xue-Feng;Yu, Hong-Shan;Wang, Dong-Ming;Liu, Ting-Qiang;Liu, Chun-Ying;An, Dong-Shan;Im, Wan-Taek;Kim, Song-Gun;Jin, Feng-Xie
    • Journal of Microbiology and Biotechnology
    • /
    • v.22 no.3
    • /
    • pp.343-351
    • /
    • 2012
  • In this paper, the kinetics of a cloned special glucosidase, named ginsenosidase type III hydrolyzing 3-O-glucoside of multi-protopanaxadiol (PPD)-type ginsenosides, were investigated. The gene (bgpA) encoding this enzyme was cloned from a Terrabacter ginsenosidimutans strain and then expressed in E. coli cells. Ginsenosidase type III was able to hydrolyze 3-O-glucoside of multi-PPD-type ginsenosides. For instance, it was able to hydrolyze the 3-O-${\beta}$-D-(1${\rightarrow}$2)-glucopyranosyl of Rb1 to gypenoside XVII, and then to further hydrolyze the 3-O-${\beta}$-D-glucopyranosyl of gypenoside XVII to gypenoside LXXV. Similarly, the enzyme could hydrolyze the glucopyranosyls linked to the 3-O-position of Rb2, Rc, Rd, Rb3, and Rg3. With a larger enzyme reaction $K_m$ value, there was a slower enzyme reaction speed; and the larger the enzyme reaction $V_{max}$ value, the faster the enzyme reaction speed was. The $K_m$ values from small to large were 3.85 mM for Rc, 4.08 mM for Rb1, 8.85 mM for Rb3, 9.09 mM for Rb2, 9.70 mM for Rg3(S), 11.4 mM for Rd and 12.9 mM for F2; and $V_{max}$ value from large to small was 23.2 mM/h for Rc, 16.6 mM/h for Rb1, 14.6 mM/h for Rb3, 14.3 mM/h for Rb2, 1.81mM/h for Rg3(S), 1.40 mM/h for Rd, and 0.41 mM/h for F2. According to the $V_{max}$ and $K_m$ values of the ginsenosidase type III, the hydrolysis speed of these substrates by the enzyme was Rc>Rb1>Rb3>Rb2>Rg3(S)>Rd>F2 in order.

Comparison of Expression Profiling of Gastric Cancer by O1igonucleotide and cDNA Microarrays (O1igonucleotide Microarray와 cDNA Microarray를 이용한 위암조직의 대단위 유전자 발현 비교)

  • Jung, Kwang-Hwa;Kim, Jung-Kyu;Noh, Ji-Heon;Eun, Jung-Woo;Bae, Hyun-Jin;Lee, Sug-Hyung;Park, Won-Sang;Yoo, Nam-Jin;Lee, Jung-Young;Nam, Suk-Woo
    • YAKHAK HOEJI
    • /
    • v.51 no.3
    • /
    • pp.179-185
    • /
    • 2007
  • Gastric cancer is one of the most common malignancies in Korea, but the predominant molecular event underlying gastric carcinogenesis remain unknown. Recently, DNA microarray technology has enabled the comprehensive analysis of gene expression level, and as such has yielded great insight into the molecular nature of cancer, However, despite the powerful approach of this techniques, the technical artifacts and/or bias in applied array platform limited the liability of resultant tens of thousand data points from microarray experiments. Therefore, we applied two different any platforms, such as olignucleotide microarray and cDNA microarray, to identify gastric cancer related large-scale molecular signature of the same human specimens. When thirty sets of matched human gastric cancer and normal tissues subjected to oligonucleotide microarray, total 623 genes were resulted as differently expressed genes in gastric cancer compared to normal tissues, and 252 genes for cDNA microarray analysis. In addition, forty three outlier genes which reflect the characteristic expression signature of gastric cancer beyond array platform and analytical protocol was recapitulated from two different expression profile. In conclusion, we were able to identify robust large-scale molecular changes in gastric cancer by applying two different platform of DNA microarray, this may facilitate to understand molecular carcinogenesis of gastric cancer.

A Comparative Analysis of Informatization Level for Agricultural Corporations and SMEs (농업법인과 중소기업의 정보화수준 비교 분석)

  • Bock, Gene;Kim, Bae-Bong;Lee, Jae-Keun
    • The Journal of Korean Institute of Communications and Information Sciences
    • /
    • v.40 no.5
    • /
    • pp.892-902
    • /
    • 2015
  • Agri-food ICT(Information and Communications Technologies) convergence has been raised as an important issue for agricultural industry competence. In this situation, this study is to enhance agricultural competitiveness and seek to development plan for agricultural corporation by diagnosing informatization level. For this purpose, this study conducted survey on informatization level of 3,019 agricultural corporations and calculated level score. And result is compared with SMEs(Small and Medium Enterprise) informatization survey, including manufacturing and service industries, conducted by Korea Technology & Information Promotion Agency for SMEs in recent agricultural corporations' growing with automation of agricultural production and improving service to customer satisfaction. Evaluation system is established to calculate informatization level score and AHP(Analytic Hierarchy Process) method was used by the experts to investigate weighting of assessment area, assessment indicators, assessment items. As a result, agricultural corporation informatization level score was 40.16 points which is lower than the benefitted organization of agri-food IT convergence modeling(43.44 points). By assessment area, the informatization level of promotional environment area was low and investment and training items were analyzed low especially so need to improve urgently. In the analysis result by organization type, agricultural company corporation's informatization level was higher than the agricultural association corporation and 'Processing and distribution' was higher than others by business type. Informatization level of agricultural corporation is 80 percent of 2013 SMEs' level(50.18 points) and 59.4 percent of a large corporation(67.64 points). In particular, big difference is occurred in investment feasibility analysis, informatization investment and education which will be need to improve.

Screening of lovastatin-producing strains by PCR using lovastatin biosynthesis genes (Lovastatin 생합성 유전자를 이용한 lovastatin 생산균주의 탐색)

  • Ko, Hee-Sun;Kim, Hyun-Soo
    • KSBB Journal
    • /
    • v.24 no.2
    • /
    • pp.163-169
    • /
    • 2009
  • Lovastatin (also known as Mevinolin, Mevacor, and Monacolin K), an inhibitor of the HMG-CoA reductase produced by Aspergillus terreus and other fungi, is used to reduce serum cholesterol levels in human beings. It is derived biosynthetically from two polyketides. One of these is a nonaketide that undergoes cyclization at a hexahydronaphthalene ring system, and the other is a simple diketide, 2-methylbutyrate. Two primer pairs were designed based on the amino acid sequences of lovastatin polyketide synthase and lovastatin diketide synthase for the PCR screening of lovastatin-producing strains. Among the seven selected strains, SJ-2 evidenced the highest level of lovastatin production in both liquid and solid cultures. Soybeans with SJ-2 were treated via 1 hour of heat shock at $30^{\circ}C$ for the mass production of lovastatin. The heat-treated soybeans were inoculated on rice bran and the koji extract was obtained after 15 days of incubation. It yielded the highest level of lovastatin production among the strains, and also evidenced 75% inhibition activity against HMG-CoA reductase. We developed an efficient PCR screening method for lovastatin-producing strains, using lovastatin biosynthesis genes.

Partial denture metal framework may harbor potentially pathogenic bacteria

  • Mengatto, Cristiane Machado;Marchini, Leonardo;de Souza Bernardes, Luciano Angelo;Gomes, Sabrina Carvalho;Silva, Alecsandro Moura;Rizzatti-Barbosa, Celia Marisa
    • The Journal of Advanced Prosthodontics
    • /
    • v.7 no.6
    • /
    • pp.468-474
    • /
    • 2015
  • PURPOSE. The aim of this study was to characterize and compare bacterial diversity on the removable partial denture (RPD) framework over time. MATERIALS AND METHODS. This descriptive pilot study included five women who were rehabilitated with free-end mandibular RPD. The biofilm on T-bar clasps were collected 1 week ($t_1$) and 4 months ($t_2$) after the RPD was inserted ($t_0$). Bacterial 16S rDNA was extracted and PCR amplified. Amplicons were cloned; clones were submitted to cycle sequencing, and sequences were compared with GenBank (98% similarity). RESULTS. A total of 180 sequences with more than 499 bp were obtained. Two phylogenetic trees with 84 ($t_1$) and 96 ($t_2$) clones represented the bacteria biofilm at the RPD. About 93% of the obtained phylotypes fell into 25 known species for $t_1$ and 17 for $t_2$, which were grouped in 5 phyla: Firmicutes ($t_1=82%$; $t_2=60%$), Actinobacteria ($t_1=5%$; $t_2=10%$), Bacteroidetes ($t_1=2%$; $t_2=6%$), Proteobacteria ($t_1=10%$; $t_2=15%$) and Fusobacteria ($t_1=1%$; $t_2=8%$). The libraries also include 3 novel phylotypes for $t_1$ and 11 for $t_2$. Library $t_2$ differs from $t_1$ (P=.004); $t_1$ is a subset of the $t_2$ (P=.052). Periodontal pathogens, such as F. nucleatum, were more prevalent in $t_2$. CONCLUSION. The biofilm composition of the RPD metal clasps changed along time after RPD wearing. The RPD framework may act as a reservoir for potentially pathogenic bacteria and the RPD wearers may benefit from regular follow-up visits and strategies on prosthesis-related oral health instructions.

Probiotic Potential of Pediococcus pentosaceus BCNU 9070 (프로바이오틱 Pediococcus pentosaceus BCNU 9070 균주)

  • Shin, Hwa-Jin;Choi, Hye-Jung;Kim, Dong-Wan;Ahn, Cheol-Soo;Lee, Young-Geun;Jeong, Young-Kee;Joo, Woo-Hong
    • Journal of Life Science
    • /
    • v.22 no.9
    • /
    • pp.1194-1200
    • /
    • 2012
  • Lactic acid bacteria are generally recognized as beneficial probiotic organisms. Recent studies revealed that the potential of probiotic strains was essentially dependent on the bacterial-binding and adhesion capabilities to gut epithelial cells and the hydrophobicity of the cell surface. In this study, we screened some indigenous lactic acid bacteria from Kimchi and selected one lactic acid bacterium as a potential probiotic based on its cell surface hydrophobicity. Analysis of the 16S rRNA gene sequences of probiotic isolates indicated that the selected isolate (BCNU 9070 strain) was a member of Pediococcus pentosaceus. P. pentosaceus BCNU 9070 showed resistance to bile acids and acidic pH. The P. pentosaceus BCNU 9070 strain also inhibited the cell growth of six food-borne pathogens including Listeria monocytogenes and Shigella sonnei. In addition, the P. pentosaceus BCNU 9070 strain expressed bile salt hydrolase activity and showed an ability to assimilate cholesterol in vitro. On the basis of these results, P. pentosaceus BCNU 9070 is considered to have probiotic potential for applications in functional foodstuffs.

Exploring Requirements of the Smart Textiles for Bio-Signal Measurement Based on Smart Watch User Sensibility (스마트워치 사용자감성에 기반한 생체신호측정용 스마트 텍스타일의 요구조건 탐색)

  • Jang, Eunji;Kim, Inhwan;Lee, Eu-Gene;Cho, Gilsoo
    • Science of Emotion and Sensibility
    • /
    • v.20 no.4
    • /
    • pp.89-100
    • /
    • 2017
  • Since smart devices are able to efficiently provide information without barriers of time and location, they are widely utilized with advent of the hyper-connected society. Especially, the smart devices have been developed in the form of wearable devices for mutual interaction between human and objects. Smart clothing, which embeds smart devices within clothes, measures and obtains a variety of bio-signals as it is in close contact with the human bodies. Conventional smart clothing generated wearers' discomfort because they were developed by simple attachment of electronic devices to clothes. Therefore, it is highly recommended to develop novel smart clothing based on smart textiles which integrate electronic devices as parts of textiles. As smart watches are currently the most available wearable devices in the market, smart watch users were selected in this study, for the purpose of investigating core needs of wearable smart device users based on the user experience and user's sensibility. Qualitative research was performed through semi-structured interview in order to obtain detailed answers about user sensibility based on smart watch user experience. After the in-depth interview, the user's sensibility was categorized into four aspects; functional, aesthetic, social, and empirical. Sensibility adjectives and key words were assigned to each aspect and their frequency was analyzed. It was the functional aspect of sensibility that the wearable device users require the most. The results of this study will be utilized as a fundamental data to develop the smart textiles required for the next generation of smart clothing which is attracting as a future wearable device.

The Robust Phylogeny of Korean Wild Boar (Sus scrofa coreanus) Using Partial D-Loop Sequence of mtDNA

  • Cho, In-Cheol;Han, Sang-Hyun;Fang, Meiying;Lee, Sung-Soo;Ko, Moon-Suck;Lee, Hang;Lim, Hyun-Tae;Yoo, Chae-Kyoung;Lee, Jun-Heon;Jeon, Jin-Tae
    • Molecules and Cells
    • /
    • v.28 no.5
    • /
    • pp.423-430
    • /
    • 2009
  • In order to elucidate the precise phylogenetic relationships of Korean wild boar (Sus scrofa coreanus), a partial mtDNA D-loop region (1,274 bp, NC_000845 nucleotide positions 16576-1236) was sequenced among 56 Korean wild boars. In total, 25 haplotypes were identified and classified into four distinct subgroups (K1 to K4) based on Bayesian phylogenetic analysis using Markov chain Monte Carlo methods. An extended analysis, adding 139 wild boars sampled worldwide, confirmed that Korean wild boars clearly belong to the Asian wild boar cluster. Unexpectedly, the Myanmarese/Thai wild boar population was detected on the same branch as Korean wild boar subgroups K3 and K4. A parsimonious median-joining network analysis including all Asian wild boar haplotypes again revealed four maternal lineages of Korean wild boars, which corresponded to the four Korean wild boar subgroups identified previously. In an additional analysis, we supplemented the Asian wild boar network with 34 Korean and Chinese domestic pig haplotypes. We found only one haplotype, C31, that was shared by Chinese wild, Chinese domestic and Korean domestic pigs. In contrast to our expectation that Korean wild boars contributed to the gene pool of Korean native pigs, these data clearly suggest that Korean native pigs would be introduced from China after domestication from Chinese wild boars.

Effect of Antibiotics and Herbicide on Shoot Regeneration from Cotyledon and Hypocotyl Explants of Chinese Cabbage (항생제와 제초제가 배추 자엽 및 배축 절편체로부터의 신초 형성에 미치는 영향)

  • Kang, Byung-Kook;Park, Young-Doo
    • Horticultural Science & Technology
    • /
    • v.19 no.1
    • /
    • pp.17-21
    • /
    • 2001
  • To develop a selection system for regenerating plants from transformed tissues, effects of four antibiotics (kanamycin, hygromycin, carbenicillin, cefotaxime) and herbicide (phosphinotricin) on shoot regeneration from cotyledon and hypocotyl explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis) were studied. For cotyledon, shoot induction was not significantly affected by kanamycin at $1mg{\cdot}L^{-1}$, but the number of shoots formed was significantly reduced at $2mg{\cdot}L^{-1}$, and no shoots were regenerated from any explants at $6mg{\cdot}L^{-1}$ or higher. Hypocotyl explants showed similar result as cotyledon. Kanamycin at $7mg{\cdot}L^{-1}$ may be adequate for selecting Chinese cabbage transformants. Hygromycin at $4mg{\cdot}L^{-1}$ or higher completely inhibited the growth and shoot regeneration of Chinese cabbage explants. Therefore, resistance gene to hygromycin may also be used as a selective marker for Chinese cabbage transformation. Carbenicillin and cefotaxime, the cephalosporin type of antibiotics, had little effect on shoot regeneration of Chinese cabbage explants. Since carbenicillin and cefotaxime have low toxicity to Chinese cabbage, they are suitable for use in tissue culture to eliminate Agrobacterium in transformation experiments after co-cultivation. Shoot regeneration from cotyledon and hypocotyl explants was significantly reduced in presence of $1mg{\cdot}L^{-1}$ phosphinotricin (PPT) and completely inhibited by $2mg{\cdot}L^{-1}$ or higher. PPT, same as antibiotics, may also be used to select transformed cells. Since Chinese cabbage is known to be recalcitrant to in vitro shoot regeneration compared to other Brassica species, even though lower levels of selectable markers result in more transformants but simultaneously allow more untransformed escapes to develop, lower levels of antibiotics and herbicides could be successfully used as a selectable marker to reduce selection pressure.

  • PDF