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Enhanced Antioxident Effect by over Expression of Tomato β-carotene Hydroxylase Gene (ChyB) Using Agrobacterium-infiltration in Tobacco Plant (Agro-infiltration을 이용한 토마토 β-carotene hydroxylase 유전자(ChyB) 과발현 및 담배식물체의 항산화 효과 증진)

  • Choi, Yoon-Jeong;Yoon, Kyung-Young;Yun, Hae-Keun;Suh, Sang-Gon;Moon, Yong-Sun
    • Horticultural Science & Technology
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    • v.29 no.3
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    • pp.267-272
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    • 2011
  • Several reports indicated that astaxanthin and zeaxanthin have more active anticancer activity than pro-vitamin A carotenes. ${\beta}$-carotene hydroxylase is a key enzyme to synthesize zeaxanthin and astaxanthin in carotenoids biosynthesis pathway. We isolated the ChyB gene encoding ${\beta}$-carotene hydroxylase from tomato leaves. The ChyB gene (1.5Kbp) fragment was cloned into the binary vector and designated to pIG121-ChyB-tom. Agrobacterium-mediated infiltration was used for transient assay in Nicotiana benthamiana. Leaf samples were collected 0, 1, 2, 3 days after infiltration (DAI). RT-PCR result showed that the expression of ${\beta}$-carotene hydroxylase transcripts was not detected in control (0DAI), but its expression was detected after 1 DPI and increased later on. When the activity of ${\beta}$-carotene hydroxylase was measured, the 1,1-diphenyl-pricryl hydrazyl (DPPH) radical scavenging activity (27%) at 2 DAI was significantly higher than that (21%) at 0 DAI. These results indicated that anti-oxidant activity dramatically increased at 2 DAI in tobacco leaves was due to over expression of tomato ${\beta}$-carotene hydroxylase. These results can be the foundation to develop tomato cultivars with high oxy-carotenoids content using the ChyB gene transformation.

Molecular Typing of Salmonella enterica serovar Typhi Strains Isolated in Busan by Pulsed-Field Gel Electrophoresis (부산지역에서 분리된 Salmonella enterica serovar Typhi균에 대한 PFGE를 이용한 Molecular typing)

  • Min, Sang-Kee;Lee, Ju-Hyun;Park, Eun-Hee;Kim, Jung-Ah;Kim, Kyu-Won
    • Journal of Life Science
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    • v.16 no.4
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    • pp.664-671
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    • 2006
  • We obtained 424 Salmonella enterica serovar Typhi isolates from sporadic cases of infection in Busan during 1996 to 2005. We investigated the trend of antimicrobial resistance and molecular typing by pulsed-field gel electrophoresis (PFGE). Of the total 424 isolates, 6 strains (1.4%) were multidrug-resistant (MDR) S. enterica serovar Typhi isolates, 2 strains (0.5%) were resistant to only nalidixic acid, and the remaining 416 strains (98.1%) were fully susceptible to the 18 antimicrobial agent. PFGE of XbaI-digested chromosomal DNA was performed on 50 sporadic S. enterica serovar Typhi isolates with the objective of investigating the extent of genetic diversity of these isolates in our region. We could find that these isolates were much more heterogeneous and at least 32 different PFGE patterns were generated according by dice coefficient, between 0.69 and 1.0. Restriction fragment patterns consisted of 13 to 18 fragments ranged in size from 20 to 630 kb. The results confirmed that PFGE would be an useful tool for investigating surveillance of sporadic or outbreak case and assessing clonality for S. enterica serovar Typhi in Busan area. Our finding will be valuable in developing rational strategies to control this pathogen and setting the basis of an effective PulseNet system in Korea.

Characterization and Cloning of a Phytase from Escherichia coli WC7. (Escherichia coli WC7가 생산하는 Phytase의 효소특성과 그 유전자의 클로닝)

  • 최원찬;오병철;김형권;강선철;오태광
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.1-7
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    • 2002
  • Phytase from Escherichia coli WC7 was purified from cell extracts and its molecular mass was estimated to be 45 kDa by SDS-PAGE. Its optimum temperature and pH for phytate hydrolysis was 6$0^{\circ}C$ and pH 5.0, respectively. The enzyme was stable up to 6$0^{\circ}C$ and over broad pH range (pH 2-12). The enzyme had higher affinity for sodium phytate than p-nitrophenylphosphate (pNPP). That is, the apparent Km value for sodium phytate and pNPP were $0.15\pm$0.02 mM and 2.82$\pm$0.05 mM, respectively. The gene encoding the phytase was cloned in E. coli XL1-Blue. Sequence analysis showed an open reading frame of 1241 Up encoding a signal peptide (22 aa) and a mature enzyme (410 aa). WC7 phytase was expressed up to 17.5 U/ml in the transformed E. coli XL1-Blue/pUEP, which was 23-fold higher than the activity from wild strain.

Analysis of Genetic Relationship of Pear (Pyrus spp.) Germplasms Using AFLP Markers (AFLP 표지를 이용한 배 유전자원의 유연관계 분석)

  • Cho, Kang-Hee;Shin, Il Sheob;Kim, Hyun Ran;Kim, Jeong-Hee;Heo, Seong;Yoo, Ki Yeol
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.444-450
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    • 2009
  • Amplified fragment length polymorphism (AFLP) marker was utilized for evaluation of genetic diversity of 60 pear germplasms. Twenty selective AFLP primer pairs generated a total of 522 polymorphic amplification products. From UPGMA (unweighted pair-group method arithmetic average) cluster analysis by using polymorphic bands, the pear germplasms were divided into four clusters by similarity index of 0.691. The first cluster (I) included European pears belonging to Pyrus communis and wild species such as P. nivalis and P. cordata. The second cluster (II) included Ussurian pea pears belonging to P. betulaefolia and P. fauriei. The third cluster (III) included pea pears belonging to P. calleryana and P. koehnei. Most of germplasms belonging to P. pyrifolia and P. ussuriensis, and interspecific hybrids were included in the fourth (IV) cluster. Therefore pear germplasms originated from East Asia were closely related to P. pyrifolia and P. ussuriensis. Similarity values among the tested pear germplasms ranged from 0.584 to 0.879, and the average similarity value was 0.686.

Mitochondrial DNA variation and phylogeography of native Mongolian goats

  • Ganbold, Onolragchaa;Lee, Seung-Hwan;Paek, Woon Kee;Munkhbayar, Munkhbaatar;Seo, Dongwon;Manjula, Prabuddha;Khujuu, Tamir;Purevee, Erdenetushig;Lee, Jun Heon
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.6
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    • pp.902-912
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    • 2020
  • Objective: Mongolia is one of a few countries that supports over 25 million goats, but genetic diversity, demographic history, and the origin of goat populations in Mongolia have not been well studied. This study was conducted to assess the genetic diversity, phylogenetic status and population structure of Mongolian native goats, as well as to discuss their origin together with other foreign breeds from different countries using hypervariable region 1 (HV1) in mtDNA. Methods: In this study, we examined the genetic diversity and phylogenetic status of Mongolian native goat populations using a 452 base-pair long fragment of HVI of mitochondrial DNA from 174 individuals representing 12 populations. In addition, 329 previously published reference sequences from different regions were included in our phylogenetic analyses. Results: Investigated native Mongolian goats displayed relatively high genetic diversities. After sequencing, we found a total of 109 polymorphic sites that defined 137 haplotypes among investigated populations. Of these, haplotype and nucleotide diversities of Mongolian goats were calculated as 0.997±0.001 and 0.0283±0.002, respectively. These haplotypes clearly clustered into four haplogroups (A, B, C, and D), with the predominance of haplogroup A (90.8%). Estimates of pairwise differences (Fst) and the analysis of molecular variance values among goat populations in Mongolia showed low genetic differentiation and weak geographical structure. In addition, Kazakh, Chinese (from Huanghuai and Leizhou), and Arabian (Turkish and Baladi breeds) goats had smaller genetic differentiation compared to Mongolian goats. Conclusion: In summary, we report novel information regarding genetic diversity, population structure, and origin of Mongolian goats. The findings obtained from this study reveal that abundant haplogroups (A to D) occur in goat populations in Mongolia, with high levels of haplotype and nucleotide diversity.

A Case Study of Mortality Caused by Viral Encephalopathy and Retinopathy (VER) in Cultured Sevenband Grouper, Epinephelus septemfasciatus during Winter (겨울철 양식 능성어의 바이러스성뇌망막증(VER) 감염사례)

  • Won, Kyoung Mi;Lee, Jeong Tae;Cho, Mi Young;Kim, Myoung Sug;Kim, Na Young;Jung, Sung Hee;Lee, Nam Sil
    • Korean Journal of Ichthyology
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    • v.29 no.3
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    • pp.157-164
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    • 2017
  • In 2015, a nervous necrosis virus (NNV) was isolated from sevenband grouper, Epinephelus septemfasciatus, maintained in land-based aquaculture system at below $12^{\circ}C$ in winter. Mortality was up to 30% in brood fish, over 4 kg of body weight. Moribund fish showed clinical sings typical of viral encephalopathy and retinopathy (VER), also called viral nervous necrosis (VNN), such as uncoordinated, corkscrew-like swimming behavior, belly-up at rest, darkening of body, cloudy eyeball and hyperinflation of the swim bladder. Aetiology of the disease was confirmed by gross observation of clinical signs, histopathology and molecular diagnosis. Histological studies revealed severe vacuolation and necrosis in the brain. Molecular diagnosis by revere transcription-polymerase chain reaction (RT-PCR) specific to batanodavirus yielded a positive result. The nucleotide sequences of the PCR-amplified fragment were 99.48~100% similar to barfin flounder nervous necrosis virus (BFNNV) genotype and most closely aligned with Pacific cod betanodavirus (PCNNV). This is the first report of natural batanodavirus, NNV infection in sevenband grouper reared in low water temperature during winter (below $12^{\circ}C$) in Korea.

A study on the Initial Nanopore Formation in the Calix Arene Based Porogen Templated Porous Thin Film (칼릭스아렌 포로젠을 이용한 다공성 박막의 초기 나노기공 형성과정에 관한 연구)

  • Kim, Do-Hun;Yim, Jin-Heong
    • Korean Chemical Engineering Research
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    • v.49 no.5
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    • pp.669-675
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    • 2011
  • Fourier Transform Infrared Spectroscopy and in-situ Position Annihilation Lifetime Spectroscopy(PALS) analysis of hybrid film, which consist of silsesquioxane(SSQ) and 4-tert-butyl calix[4]arene-O,O',O",O'"-tetraacetic acid tetraethyl ester(CA[4]) have been investigated in order to understand initial formation of nanopore in the next generation porous low-k dielectrics(k < 2.0). SSQ/CA[4] can provide effective homogeneous thin film having porous structure. The porogen decomposition behavior were completely different in the two kinds of SSQ/CA[4] based hybrid film (i.e. SSQ/CA[4] 10 and SSQ/CA[4] 20%). Relatively small pores(1.5 nm) come from dispersion of uni-molecular CA[4] in the SSQ matrix have been generated at $300^{\circ}C$, while mesopores(2.5~3.0 nm) induced from self assembled CA[4] have been generated at $250^{\circ}C$. It might be due to highly interconnected structure of SSQ/CA[4] 20% hybrid thin film resulting in facile evacuating of decomposed fragment of CA[4] molecule.

Polymorphisms of 5,10-Methylenetetrahydrofolate Reductase (MTHFR C677T and A1298C) Gene in Recurrent Spontaneous Abortion (5,10-Methylenetetrahydrofolate Reductase (MTHFR C677T와 A1298C) 유전자 돌연변이의 반복자연유산 관련성 연구)

  • Kim, Nam-Keun;Nam, Yoon-Sung;Lee, Su-Man;Kim, Sun-Hee;Shin, Seung-Joo;Chang, Sung-Woon;Kim, Se-Hyun;Cha, Kwang-Yul;Oh, Do-Yeun
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.3
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    • pp.215-222
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    • 2002
  • Objective : Previous studies have suggested that hyperhomocysteinemia and methylenetetrahydrofolate reductase (MTHFR C677T) mutations are associated with increased risk of recurrent spontaneous abortion (RSA). Recently, a second site polymorphism in MTHFR, 1298A-->C, which changes a glutamic acid into an alanine residue, was shown to be associated with a decreased enzyme activity. We tested whether the variant alleles of MTHFR C677T and A1298C are risk factor (biomarker) for RSA. Materials and Methods: We analyzed DNA from a case-control study in the Korean DNA was extracted from blood samples of 118 patients with RSA and 123 healthy fertile patients as the controls. MTHFR variant alleles were determined by a PCR-restriction fragment length polymorphism assay. Results: We found no evidence for an association between 677TT genotype and risk of RSA (OR=1.95, 95% CI=$0.84{\sim}4.50$, p=0.12). However, the MTHFR 1298AC (OR=0.36, 95% CI=$0.20{\sim}0.63$, p=0.0004) and 1298AC+CC (OR=0.35, 95% CI=$0.20{\sim}0.61$, p=0.0002) genotypes were lower among 118 RSA cases compared with 123 controls, conferring a 2.8-fold decrease in risk of RSA, respectively. Moreover, the combined genotypes of MTHFR 677CC/1298AC (OR=0.30, 95% CI=$0.10{\sim}0.88$, p=0.029) and 677CT/1298AC (OR=0.77, 95% CI=$0.60{\sim}0.99$, p=0.043) also showed significantly lower risk than those with MTHFR 677CC/1298AA type. Conclusion: MTHFR 1298AC, MTHFR 677CC/1298AC and 677CT/1298AC genotypes may represent genetic markers for the protection of RSA at least in Korean women.

The Study of 5,10-Methylenetetrahydrofolate Reductase Variation (MTHFR C677T) in Infertile Females with Polycystic Ovarian Syndrome (PCOS) in Korea (한국인 다낭성 난포증후군 환자에서 5,10-Methylenetetrahydrofolate Reductase의 677번 유전자 다형성에 관한 연구)

  • Lee, Kyo-Won;Jeong, Yu-Mi;Lee, Sook-Hwan;Yoon, Tae-Ki;Kwak, In-Pyung;Yoon, Seon-Woong;Choi, Joong-Sub;Kim, Kye-Hyun;Han, Jong-Sul;Kim, Sung-Do;Kim, Nam-Keun;Cha, Kwang-Yul;Baek, Kwang-Hyun;Lee, Su-Man
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.3
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    • pp.217-222
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    • 2003
  • Objective: To investigate the association of genetic background between MTHFR C677T genotype and infertile females with polycystic ovarian syndrome. Materials and Methods: We compared 86 infertile females with polycystic ovarian syndrome (PCOS) with 100 healthy fertile females with one or more offspring. Pyrosequencing analysis for MTHFR C677T variation was performed on polymerase chain reaction (PCR) product of study group. To validate pyrosequencing data of C677T variation for randomly selected 50 samples, we compared the pyrosequencing result with the PCR-RFLP (Restriction Fragment Length Polymorphism) result of MTHFR C677T genotype. Results: The prevalence of the C677T mutant homozygous (TT) was significantly lower (p=0.0085) in females with PCOS (8.14%) than in fertile females (21.00%). MTHFR 677 TT genotype had a decreased risk (3.7-fold) of PCOS compared with wild type (MTHFR 677 CC). Conclusion: Our data support a role for MTHFR mutant homozygous (677 TT) genotype in reducing risk in Korean infertile females with Polycystic ovarian syndrome.

Binding of the His-tagged Tail Protein J of Bacteriophage Lambda with Escherichia coli K-12 (히스티딘으로 표지된 람다 박테리오파아지 꼬리 단백질 J와 대장균 K-12와의 결합)

  • Shin, Hae Ja
    • Journal of Life Science
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    • v.28 no.1
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    • pp.78-82
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    • 2018
  • Detection of pathogenic microorganisms takes several days by conventional methods. It is necessary to assess microorganisms in a timely manner to reduce the risk of spreading infection. For this purpose, bacteriophages are chosen for use as a biosensing tool due to their host specificity, wide abundance, and safety. However, their lytic cycle limits their efficacy as biosensors. Phage proteins involved in binding to bacteria could be a robust alternative in resolving this drawback. Here, a fragment of tail protein J (residues 784 to 1,132) of phage lambda fused with 6X His-tag (6HN-J) at its N-terminus was cloned, overexpressed, purified, and characterized for its binding with microorganisms. The purified protein demonstrated a size of about 38 kDa in sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) and bound with anti-His monoclonal antibodies. It bound specifically to Escherichia coli K-12, and not Salmonella typhimurium, Bacillus subtilis, or Pseudomonas aeruginosa in dot blotting. Binding of the protein to E. coli K-12 inhibited about 50% of the in vivo adsorption of the phage lambda to host cells at a concentration of $1{\mu}g/ml$ 6HN-J protein and almost 100% at $25{\mu}g/ml$ 6HN-J. The results suggest that a fusion viral protein could be utilized as a biosensing element (e.g., protein chips) for detecting microorganisms in real time.