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Characterization of Exolytic GH50A β-Agarase and GH117A α-NABH Involved in Agarose Saccharification of Cellvibrio sp. KY-GH-1 and Possible Application to Mass Production of NA2 and L-AHG (Cellvibrio sp. KY-GH-1의 아가로오스 당화 관련 엑소형 GH50A β-아가레이즈와 GH117A α-NABH의 특성 및 NA2와 L-AHG 양산에의 적용 가능성)

  • Jang, Won Young;Lee, Hee Kyoung;Kim, Young Ho
    • Journal of Life Science
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    • v.31 no.3
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    • pp.356-365
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    • 2021
  • Recently, we sequenced the entire genome of a freshwater agar-degrading bacterium Cellvibrio sp. KY-GH-1 (KCTC13629BP) to explore genetic information encoding agarases that hydrolyze agarose into monomers 3,6-anhydro-L-galactose (L-AHG) and D-galactose. The KY-GH-1 strain appeared to possess nine β-agarase genes and two α-neoagarobiose hydrolase (α-NABH) genes in a 77-kb agarase gene cluster. Based on these genetic information, the KY-GH-1 strain-caused agarose degradation into L-AHG and D-galactose was predicted to be initiated by both endolytic GH16 and GH86 β-agarases to generate NAOS (NA4/NA6/NA8), and further processed by exolytic GH50 β-agarases to generate NA2, and then terminated by GH117 α-NABHs which degrade NA2 into L-AHG and D-galactose. More recently, by employing E. coli expression system with pET-30a vector we obtained three recombinant His-tagged GH50 family β-agarases (GH50A, GH50B, and GH50C) derived from Cellvibrio sp. KY-GH-1 to compare their enzymatic properties. GH50A β-agarase turned out to have the highest exolytic β-agarase activity among the three GH50 isozymes, catalyzing efficient NA2 production from the substrate (agarose, NAOS or AOS). Additionally, we determined that GH117A α-NABH, but not GH117B α-NABH, could potently degrade NA2 into L-AHG and D-galactose. Sequentially, we examined the enzymatic characteristics of GH50A β-agarase and GH117A α-NABH, and assessed their efficiency for NA2 production from agarose and for production of L-AHG and D-galactose from NA2, respectively. In this review, we describe the benefits of recombinant GH50A β-agarase and GH117A α-NABH originated from Cellvibrio sp. KY-GH-1, which may be useful for the enzymatic hydrolysis of agarose for mass production of L-AHG and D-galactose.

A Study of Microbial Contamination in Fresh-Cut and Ready-to-Eat Foods Purchased from Online Markets (온라인 판매 신선편의식품 및 즉석섭취식품의 미생물 오염도 연구)

  • Hye-Sun Hwang;Jae-Hoon Jeong;Young-Hee Kwon;Ye-Jee Byun;Ji-Young Park;Ho-Cheol Yun
    • Journal of Food Hygiene and Safety
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    • v.39 no.4
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    • pp.335-342
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    • 2024
  • This study aimed to examine the delivery conditions and microbial contamination of fresh-cut and ready-to-eat foods purchased from online markets between February and November 2023. Upon arrival, the average surface temperature of the products was 11.3℃. In the fresh-cut foods, the average number of total aerobic bacteria and coliforms was 4.5 log colony-forming units (CFU)/g and 1.2 log CFU/g, respectively, whereas in the ready-to-eat foods, these values were 10.6 log CFU/g and 1.2 log CFU/g, respectively. Pathogens, such as Staphylococcus aureus, Salmonella spp., Clostridium perfringens, Listeria monocytogenes, and pathogenic Escherichia coli were absent from all samples. Bacillus cereus was found in 2.7% of the fresh-cut foods and 0.9% of the ready-to-eat foods, with contamination levels averaging 0.05 log CFU/g and 0.01 log CFU/g, respectively. In the four samples in which B. cereus was detected, genetic testing of the six toxin genes produced by B. cereus revealed the presence of at least one enterotoxin gene, excluding the emetic toxin. L. monocytogenes was absent from ready-to-eat foods but was detected in 0.9% of fresh-cut foods. Analysis of the isolated L. monocytogenes confirmed the presence of six pathogenicity-related genes, including iap, indicating the potential risk of foodborne diseases.

Studies on the Nutritional Effects of Locally Produced Dried Formula Milk and Fermented Milk -2. Effect of Feeding Dried Formula Milk and Fermented Milk on the Growing Performance, Protein Utilizability and Intestinal Microbial Changes of Albino Rats (국산(國産) 조제분유(調製粉乳)와 발효유(醱酵乳)의 영양효과(營養效果)에 관한 연구(硏究) - 2. 유제품(乳製品)이 성장기(成長期) 흰쥐에 미치는 영양효과(營養效果) 및 장내미생물(腸內微生物)의 변화(變化)에 관한 연구(硏究) -)

  • Paik, Jeong-J.;Han, In-K.
    • Journal of Nutrition and Health
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    • v.9 no.1
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    • pp.92-98
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    • 1976
  • The present studies were carried out to observe the nutritional effects of three kinds of locally produced dried infantile formula milk (DFM) and one fermented milk (FM). A feeding trial with 60 male growing albino rats weighing $60{\sim}70$ grams was conducted during 6 weeks to compare the nutritive values and protein qualities of three DFM and a FM. The diet treatments consisted of 100% control diet, 70% control diet plus 30% DFM-A, 70% control diet plus 30% DFM-B, 70% control diet plus 30% DFM-C, 100% control diet with FM and 70% control diet plus 30% DFM-B with FM. The items investigated were body weight gain, feed intake, feed efficiency ratio (FER), various organ weights, protein efficiency ratio (PER), digestibility of nutrients, biological value, utilizability of protein and intestinal microbial changes of albino rats. The results obtained are summarized as follows; 1. Although there was no statistical significance, rats fed diets containing DFMs and FM gained faster than the rats fed control diet. The best growth rate was obtained with the DFM-A and DFM-C groups. In spite of the lower protein contents of the three DFM diets than the control diet, the growth rate of albino rats fed the DFM diets was somewhat improved than rats fed control diet. 2. No statistical significance was found in feed consumption but the trend was that the feed intake of control group was higher than those of the DFM diet group. 3. Feed efficiency was inproved significantly (p<0.01) by feeding DFMs as compared with control diet. DFM-A group showed the best FER, although no statistical significance was found. 4. Rats fed the DFM diets showed significantly (p<0.01) higher PER as compared with those of the control group. But no difference was found anions DFM groups. The significant improvement (p<0.01) of PER due also to the feeding of FM was seemed to be brought about by the beneficial effect of FM. 5. The present data revealed that feeding DFM and FM didn't affect the weights of various organs of rats. 6. The protein digestibility of experimental diets was similar to each other. Although no statistical significance was found among treatments, the DFMs and FM surely tended to improve the biological value and utilizability of protein. 7. Microbial study indicated that among intestinal flora FM fed group, there were more Lactic acid bacteria than E. coli. From the experimental results described above, it may be concluded that the nutritive effects of three kinds of locally produced DFMs are much alike and the growth rate of growing albino rats can be improved by feeding either DFM or FM due to their beneficial effects on the feed efficiency and protein utilization.

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Effect of Supplementary Actinomycetes (Nocardia sp. CS682) Ferment on the Laying Performance, Blood Parameters, Immunoglobulin and Small Intestinal Microflora Contents in Laying Hens (방선균(Nocardia sp. CS682) 발효물의 급여가 산란계의 생산성, 혈액성상, 면역글로불린 및 소장내 미생물 함량에 미치는 영향)

  • Rhee, Ah-Reum;Shin, Dong-Hun;Kim, Chan-Ho;Jung, Byoung-Yun;Yoo, Jin-Chul;Hong, Young-Ho;Paik, In-Kee
    • Korean Journal of Poultry Science
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    • v.38 no.1
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    • pp.59-69
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    • 2011
  • This study was conducted to investigate the effects of dietary supplementation of CS682, a fermentation product of Actinomycetae(Nocardia sp. CS682), and its commercial product DSC682$^{(R)}$ on the performance, blood parameters, intestinal microflora, and immune response in laying hens. Hy-Line Brown$^{(R)}$ laying hens were housed in two bird cages. Feeding trial lasted 5 wk under 16.5 h:7.5 h(L:D) lighting regimen. In Exp.1, a total of 480 birds of 86 wk old were assigned to four dietary treatments: Control, Antibiotics (6 ppm avilamycin), CS682-0.1 (CS682 0.1%) and CS682-1.0 (CS682 1.0% supplementation). Each treatment was replicated five times with 24 birds (or 12 cages) per replication. In Exp. 2, a total of 1,000 birds of 26 wk old were assigned to five dietary treatments: Control, Antibiotics (6 ppm avilamycin), DCS682-0.05 (DCS682 0.05%), DCS682-0.1 (DCS682 0.1%), DCS682-0.2 (DCS682 0.2% supplementation). Each treatment was replicated five times with 40 birds (or 20 cages) per replication. In Exp. 1, there were no significant differences among treatments in egg production, egg weight, broken & soft egg production, feed intake, and feed conversion ratio. Also, there were no significant differences among treatments in eggshell thickness, eggshell color and Haugh unit. However, eggshell strength was significantly (p<0.05) greater in CS682 and Antibiotics treatments than Control, and egg yolk color was significantly (p<0.05) higher in CS682-1.0 than Control. In Exp. 2, feed intake was significantly (p<0.05) lower in DSC682-0.05 than Control. Lightness(L) of Hunter Lab color of eggshell of DCS and Antibiotics treatments was significantly (p<0.05) lower than Control. Egg yolk color of DCS 0.1 and 0.2 treatments was significantly (p<0.05) higher than Control. Haugh unit increased significantly (p<0.05) in Antibiotics and DCS682-0.1 treatments. The immunoglobulin levels of plasma (IgG and IgA) and eggyolk (IgY) were not significantly affected by treatments. Antibiotics and CS682 or DCS682 treatments significantly (p<0.05 or 0.01) influenced some of the erythrocytes and leukocytes parameters in blood. In Exp.1, mean corpuscular volume (MCV) decreased by CS682 treatments and mean corpuscular hemoglobin (MCH) was highest in Antibiotics treatments. In Exp.2, the level of monocyte (MO) decreased in DCS682-0.10 and 0.20 treatments. The cfu of C. perfringens and S. typhimurium in small intestinal content were highest in Control and lowest in Antibiotics in both experiments. In Exp. 2, DSC682-0.05 and -0.1 treatments were highest and Antibiotic treatment was lowest in Lactobacilli spp. The results of the present layer experiments indicated that supplementation of 0.1~0.2% CS682 or DCS682 may increase eggshell strength, color of eggshell and eggyolk, Haugh unit, and control harmful intestinal microbes.

Microbial Hazard Analysis of Astragalus membranaceus Bunge for the Good Agricultural Practices (농산물우수관리를 위한 황기(Astragalus membranaceus Bunge)의 미생물학적 위해요소 분석)

  • Kim, Yeon Rok;Lee, Kyoung Ah;Kim, Se-Ri;Kim, Won-Il;Ryu, Song Hee;Ryu, Jae-gee;Kim, Hwang-Yong
    • Journal of Food Hygiene and Safety
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    • v.29 no.3
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    • pp.181-188
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    • 2014
  • The objective of this study was to analyze the microbiological hazards of Astragalus membranaceus Bunge on the post-harvest processing. Samples from processing equipments (cleaner, water, cart, table, tray and packaging machine), personal hygiene (hand) and harvested crops (before washing, after washing, after sorting, and after drying) were collected from four farms (A, B, C, and D) located in Chungchengbuk-do, Korea. The samples were analyzed for sanitary indication bacteria and pathogenic bacteria. First, total aerobic bacteria and coliform in processing facilities were detected at the levels of 0.93~4.86 and 0.33~2.28 log CFU/$100cm^2$ and/mL respectively. In particular, microbial contamination in hand (5.43~6.11 and 2.52~4.12 log CFU/Hand) showed higher than processing equipments. Among the pathogenic bacteria, Bacillus cereus was detected at the levels of 0.33~2.41 log CFU/$100cm^2$, 1.48~3.27 log CFU/Hand and 0.67~3.65 log CFU/g in equipments, hands, and plants and Staphylococcus aureus were detected in cleaner, table, hand and harvested crops (before washing and after sorting) by qualitative test. Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella spp. were not detected. These results indicated that personal hygiene and processing equipments should be managed to reduce the microbial contamination of A. membranaceus Bunge. Therefore, management system such as good agricultural practices (GAP) criteria is needed for hygienic agricultural products.

Local Shape Analysis of the Hippocampus using Hierarchical Level-of-Detail Representations (계층적 Level-of-Detail 표현을 이용한 해마의 국부적인 형상 분석)

  • Kim Jeong-Sik;Choi Soo-Mi;Choi Yoo-Ju;Kim Myoung-Hee
    • The KIPS Transactions:PartA
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    • v.11A no.7 s.91
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    • pp.555-562
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    • 2004
  • Both global volume reduction and local shape changes of hippocampus within the brain indicate their abnormal neurological states. Hippocampal shape analysis consists of two main steps. First, construct a hippocampal shape representation model ; second, compute a shape similarity from this representation. This paper proposes a novel method for the analysis of hippocampal shape using integrated Octree-based representation, containing meshes, voxels, and skeletons. First of all, we create multi-level meshes by applying the Marching Cube algorithm to the hippocampal region segmented from MR images. This model is converted to intermediate binary voxel representation. And we extract the 3D skeleton from these voxels using the slice-based skeletonization method. Then, in order to acquire multiresolutional shape representation, we store hierarchically the meshes, voxels, skeletons comprised in nodes of the Octree, and we extract the sample meshes using the ray-tracing based mesh sampling technique. Finally, as a similarity measure between the shapes, we compute $L_2$ Norm and Hausdorff distance for each sam-pled mesh pair by shooting the rays fired from the extracted skeleton. As we use a mouse picking interface for analyzing a local shape inter-actively, we provide an interaction and multiresolution based analysis for the local shape changes. In this paper, our experiment shows that our approach is robust to the rotation and the scale, especially effective to discriminate the changes between local shapes of hippocampus and more-over to increase the speed of analysis without degrading accuracy by using a hierarchical level-of-detail approach.

Studies on the Synthesis of Mannich Bases of Hexachlorophene and their Antimicrobial Activities (Hexachlorophene의 Mannich Bases 합성 및 항미생물작용에 관한 연구)

  • Kim, Jong-Ho;Bae, Moo;Lee, Gye-Jun
    • Microbiology and Biotechnology Letters
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    • v.1 no.1
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    • pp.43-50
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    • 1973
  • Thirty-three Mannich bases of 2,2'-methylene bis (3,4, 6-trichlorophenoxy-acetic acid) were synthesized hexachlorophene as potential antimicrobial agents and tested against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Trichophyton rubrum, Microsporum gypseum, Epidermophyton floccosum, Aspergillus niger and Aspergillus oryzae in vitro. It was found that: 1) 2,2'-Methylene bis [${\alpha}$-(3, 4, 6-trichlorophenoxy)-${\beta}$- (N,N -diethylamino) propionic acid] and 2,2'-methylene bis [${\alpha}$-(3,4,6-trichlorophenoxy)-${\beta}$-(N, N-dimethynlamo) propionoic acid] were active against Staphylococcus aureus and Bacillus subtilis at the concn. of 1 $\mu\textrm{g}$/$m\ell$ respectively; 2) 2,2'-Methylene bis [${\alpha}$-(3,4,6-trichlorophenoxy)-${\beta}$-(m-hydroxy-p-carboxyphenylamino) propionic acid] and 2,2'-methylene bis [${\alpha}$-(3, 4, 6-trichlorophenoxy)-${\beta}$-(cyclohexylamino) propionic acid] were active against Trichophyton-rubrum at the concn. of 2 $\mu\textrm{g}$/$m\ell$ respectively; 3) 2,2'-Methylene bis [${\alpha}$-3,4,6-trichlorophenoxy)-${\beta}$-(m-hydroxy-p-carboxyphenyl-amino) propionic acid] and 2,2'-methylene bis [${\alpha}$-(3,4,6-trcihlorophenoxy)-${\beta}$-(piperidino) propionic acid] were active against Microsporum gypseum at the concn. of 2 $\mu\textrm{g}$/$m\ell$ respectively; 4) 2,2'-Methylene bis [${\alpha}$-(3,4,6-trichlorophenoxy)-${\beta}$-(m-hydroxyphenylamino) propionic acid], 2,2'-methylene bis [${\alpha}$-(3, 4,6-trichlorophenoxy)-${\beta}$-(m-hydroxy-p-carboxy phenylamino) propionic acid], 2,2'-methylene bis [${\alpha}$-(3,4,6-trichlorophenoxy)-${\beta}$-(o-chlorophenylamino) propionic acid], 2,2'-methylene bis [${\alpha}$-(3,4,6-trichlorophenoxy)-${\beta}$-(o-chloro-p-nitrophenylamino) propionic acid], 2,2'-methylene bis [${\alpha}$- (3,4,6-trichlorophenoxy)-${\beta}$-(methylamino) propionic acid], 2,2'-methylene bis [${\alpha}$-(3,4,6-trichlorophenoxy)-${\beta}$-(hydroxylamino) propionic acid], 2,2'-methylene bis [${\alpha}$-(3,4,6-trichlorophenoxy)-${\beta}$-(cyclohexylamino) propionic acid], 2,2'-methylene bis [${\alpha}$-(3,4,6-trichlorphenoxy)-${\beta}$-(morpholino) propionic acid], 2,2'-methylene bis [${\alpha}$-(3,4,6-trichlorophenoxy)-${\beta}$-(p-sulfophenylamino) propionic acid] and 2,2'-methylene bis [${\alpha}$-(3,4,6-trichlorophenoxy)-${\beta}$-(4-sulfu-l-nayphthlamino) aoi!c rppioncd (were active against Epidermophyton floccosum at the concn. of 1 $\mu\textrm{g}$/$m\ell$ respectively; 5) 2,2'-Methlene bis [${\alpha}$-(3,4,6-trichlorophenoxy)-${\beta}$-(m-hydroxyphenylamino) propionic acid], 2,2'-methylene bis (a-(3,4,6-trichlorophenoxy)-${\beta}$-(m-hydroxy-p-carboxyphenylamino) propionic acid], 2,2'-methylene bis [${\alpha}$-(3,4,6-trichlorophenoxy)-${\beta}$-(p-methylphenylamino) propionic acid] and 2,2'-methylene bis [${\alpha}$-(3,4,6-trichlorophenoxy)-${\beta}$-(hydroxylamino) propionic acid] were active against Aspergillus niger and Aspergillus oryzae at the concn. of 1 $\mu\textrm{g}$/$m\ell$ respectively.

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Effect of Gas Mixture Ratio of Modified Atmosphere Packaging on Quality of Chicken Breast (CO2와 N2 혼합 비율에 따른 포장 닭고기 가슴육의 냉장 저장 중 품질 특성)

  • Chae, Hyun-Seok;Na, Jae-Cheon;Choi, Hee-Cheol;Kim, Min-Ji;Bang, Han-Tae;Kang, Hwan-Ku;Kim, Dong-Wook;Suh, Ok-Seok;Ham, Jun-Sang;Jang, Ae-Ra
    • Food Science of Animal Resources
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    • v.31 no.1
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    • pp.100-106
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    • 2011
  • The physicochemical characteristics of chicken breast were determined to identify the optimal ratio of $CO_2$ and $N_2$ to maintain chicken breast quality during cold storage for 6 d. The mixing ratios of $CO_2$ and $N_2$ were 20:80, 40:60, 60:40, and 80:20, respectively. The pH of the chicken breast packed with 80% $CO_2$ and 20% $N_2$ was lower than that of the control on day 1 (p<0.05). The lightness ($L^*$) of the breast increased with increasing $CO_2$ during storage (p<0.05), whereas no difference was found for redness ($a^*$) and yellowness ($b^*$). A lower volatile basic nitrogen level was found for chicken breasts exposed to higher $CO_2$ levels. Furthermore, lipid oxidation of the chicken breast packed with $CO_2$ decreased with increasing $CO_2$ level, and 40% $CO_2$ significantly reduced 2-thiobarbituric acid reactive substances (TBARS) values on days 1 and 6. The total number of microbes was reduced in chicken breast exposed to more than 40% $CO_2$ during storage days 3 and 6 (p<0.05); however, Escherichia coli was not affected by $CO_2$ level. Coliforms of chicken breast were reduced in the 40% $CO_2$ level on storage day 3. Moreover, tray-packed chicken breast exposed to 40% $CO_2$ did not collapse. These results suggest that 40% $CO_2$ and 60% $N_2$ were the optimal conditions for packaging chicken breasts during cold storage.

Evaluation on Microbiological Contamination Level of Raw Beef from Retail Markets in Seoul, Korea (서울지역 식육판매점의 우육에 대한 미생물학적 오염도 평가)

  • Ko, Eun-Kyung;Heo, Eun Jeong;Kim, Young Jo;Park, Hyun Jung;Wi, Seong-Hwan;Moon, Jin San
    • Food Science of Animal Resources
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    • v.33 no.3
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    • pp.403-410
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    • 2013
  • This study was performed to evaluate the microbiological contamination level of raw beef from retail markets in Seoul, Korea. The sampling and laboratory test were performed according to the procedure of "Standard for processing and ingredients specification of livestock product" and "Korean food code". Enterotoxin of Staphylococcus aureus isolates were detected using VIDAS$^{(R)}$ and PCR-based methods. Listeria monocytogenes serotyping and genotyping were carried out using Listeria antisera and L. monocytogenes Fingerprinting kit, respectively. A total of 48 samples were collected from 16 retail markets (butcher's shop: 5, department store: 6, supermarket: 5) in 2011. The level of total bacteria counts in the butcher's shop, department store and supermarket were $4.4{\times}10^3$ CFU/g, $3.9{\times}10^5$ CFU/g and $1.0{\times}10^4$ CFU/g, respectively. The concentrations of Escherichia coli of these three retail markets were $6.4{\times}10$ CFU/g, 7.6 CFU/g and $2.0{\times}10$ CFU/g, respectively. Salmonella species was not detected on all samples. However, S. aureus was isolated in the 3 samples (6.25%) from each type of three retail markets. L. monocytogenes was isolated in the 4 samples (8.3%) from department stores. The level of contamination of these foodborne bacteria was less than 100 CFU/g. The enterotoxin-encoding genes of S. aureus isolates were sea, seh, sei and sep gene. The gene similarity of L. monocytogenes isolated from two retail markets by Rep-PCR showed 57.8-98.1% and 68.1-98.1%, respectively. These results suggest that the HACCP guideline for environmental control in slaughterhouse and retail markets should be provided to prevent cross contamination and manage foodborne pathogens such as L. monocytogenes and S. aureus.

Construction of Retrovirus Vector System for the Regulation of Recombinant hTPO Gene Expression (재조합 hTPO 유전자의 발현 조절을 위한 Retrovirus Vector System의 구축)

  • Kwon, Mo-Sun;Koo, Bon-Chul;Kim, Do-Hyang;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.161-167
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    • 2007
  • In this study, we constructed and tested retrovirus vectors designed to express the human thrombopoietin gene under the control of the tetracycline-inducible promoters. To increase the hTPO gene expression at him-on state, WPRE sequence was also introduced into retrovirus vector at downstream region of either the hTPO gene or the sequence encoding reverse tetracycline-controlled transactivator (rtTA). Primary culture cells (PFF, porcine fetal fibroblast; CEF, chicken embryonic fibroblast) infected with the recombinant retrovirus were cultured in the medium supplemented with or without doxycycline for 48hr, and induction efficiency was measured by comparing the hTPO gene expression level using RT-PCR, western blot and ELISA. Higher hPTO expression and tighter expression control were observed from the vector in which the WPRE sequence was placed at downstream of the hTPO (in CEF) or rtTA(in PFF) gene. This resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.