• Journal of Animal Science and Technology
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• 제50권6호
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• pp.793-798
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• 2008

정액 내 세균오염 종류와 검출 빈도를 조사한 결과는 Table 1에서 보는 바와 같다. 돼지 정액 내 오염세균 중 검출빈도는 Staphylo- coccus genus, Proteus genus, Bacillus genus, Pasteulla genus, Acinetobacte genus 및 Serratia genus 등의 순으로 나타났다. 동정된 세균 중 33종의 세균에 대하여 디스크법을 이용하여 8종의 항생제에 대한 감수성을 조사 한 결과는 Table 2와 3에서 보는 바와 같다. 검출된 세균의 항생제 감수성을 조사한 결과 항생제 감수성이 높은 항생제는 Amikacin, Polymyxin B, Neomycin, Streptomycin, Kanamycin, Tetracycline, Erythromycin 및 Penicillin순 이었다. Enterococcus faecium와 Streptococcus uberis는 Streptomycin을 제외한 모든 항생제에 대하여 저항성이 있는 것으로 조사되었다.

• International Journal of Industrial Entomology and Biomaterials
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• 제2권1호
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• pp.19-26
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• 2001

Transformed Spodoptera frugiperda (Sf9) cells expressing baculovirus very late factor (VLF-1) were constructed by using Autograha nuclear polyhedrosis virus (AcNPV) immediate earthy gene (ie1). Neomycin-resistance gene as a selectable marker was introduced under the control of AcNPV ie1 promoter, and Bombyx mori nuclear polyhedrosis (BmNPV-K1) vlf-1 gene was introduced under the control of the Drosophila heat shock protein gene (hspr70) promoter to yield dual expression plasmid with two independent transcription units. It was transfected into Sf9 cells and cell clones expressing vlf-1 were selected by G4l8 treatment. Genomic DNA from transformed cells was isolated and integration of AcNPV iel harboring vlf-1 was confirmed by PCR using AcNPV iel-specific primers and Southern blot analysis. The transformed cells expressing VLF-1 in an infection-independent manner expressed foreign gene product of recombinant baculovirus in the earlier stage of infection compared with control Sf9 cells. These results suggest the possible to develop highly efficient transformed insect cells for baculovirus expression vector system.

• Asian-Australasian Journal of Animal Sciences
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• 제29권4호
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• pp.564-570
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• 2016

The Sal-like 1 gene (Sall1) is essential for kidney development, and mutations in this gene result in abnormalities in the kidneys. Mice lacking Sall1 show agenesis or severe dysgenesis of the kidneys. In a recent study, blastocyst complementation was used to develop mice and pigs with exogenic organs. In the present study, transcription activator-like effector nuclease (TALEN)-mediated homologous recombination was used to produce Sall1-knockout porcine fibroblasts for developing knockout pigs. The vector targeting the Sall1 locus included a 5.5-kb 5' arm, 1.8-kb 3' arm, and a neomycin resistance gene as a positive selection marker. The knockout vector and TALEN were introduced into porcine fibroblasts by electroporation. Antibiotic selection was performed over 11 days by using $300{\mu}g/mL$ G418. DNA of cells from G418-resistant colonies was amplified using polymerase chain reaction (PCR) to confirm the presence of fragments corresponding to the 3' and 5' arms of Sall1. Further, mono- and bi-allelic knockout cells were isolated and analyzed using PCR-restriction fragment length polymorphism. The results of our study indicated that TALEN-mediated homologous recombination induced bi-allelic knockout of the endogenous gene.

• 대한수의학회지
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• 제47권1호
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• pp.33-41
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• 2007

Streptococcus spp. comprising Streptococcus (S.) uberis S. dysgalactiae strains is major causeof bovine mastitis from particularly well-managed or low somatic cell count herds that have successfullycontrolled contagious pathogens. In this study, antimicrobial susceptibility and genetic characteristics of S.uberis isolated from clinical or subclinical mastitis milk at 2003 were investigated. Eighty seven isolatesof Streptococus spp. were identified by the conventional biochemical methods. The antimicrobialsusceptibility by disk diffusion method was determined for 46 S. uberis, 11 S. bovis, 10 S. oralis, 6 S.uberis and 14 other Streptococcus spp.. Overall, the tested strains were susceptible to tetracycline (11.5%),amikacin (14.9%), streptomycin (16.1%), neomycin (26.4%), kanamycin (35.6%), gentamicin (65.2%),oxacillin (70.1%), ampicillin (75.9%), chloramphenicol (78.2%), and cephalothin (97.7%). Additionally, S.uberis strains were susceptible to pencillin G (97.8%), but resistant to erythromycin (76.0%) by minimalinhibitory concentration test. The multiple-drug resistance rate of isolated bacteria to 4 more thanamplification fingerprinting patterns amplifed with primer 8.6d showed that 3 to 8 number of distinguishableDNA fragments ranged from 180 bp to 1,20 bp. Thirty seven isolates of S. uberis strains were subtypedinto 8 distinct patterns. Each subtype revealed a typical pattern of antimicrobial susceptibilities. Thesefindings demonstrate that S. uberis isolates were mastitis pathogens of diverse serotypes, and oftenencountered the diverse resistant patterns.

• 한국동물위생학회지
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• 제33권1호
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• pp.59-66
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• 2010

This study was carried out to investigate the characters of Listeria monocytogenes isolated from food, animal feces, dry cattle food, and the environment in Seoul and Kyonggi province during the period from 1998 to 2003. Serotyping of 70 L. monocytogenes isolates was performed according to the manufacturer's instruction. Minimum inhibitory concentrations (MICs) were determined by the microdilution method according to the guidelines of the Clinical and Laboratory Standards Institute. All the isolates were tested against 20 antimicrobial agents. The serotypes of the 70 L. monocytogenes isolates were 1/2c (62.8%), 1/2a (20%) and 1/2b (17.2%). Of the 70 L. monocytogenes isolates, 67.1%, 57.1%, 11.4%, 5.7%, 2.8%, 1.4% and 1.4% were resistant to tetracycline (Te), minocycline (Mi), norfloxacin (Nor), ciprofloxacin (Cip), neomycin (N), chloramphenicol (C) and cephalothin (Cf), respectively. However, all isolates were 100% sensitive to antibiotics such as amikacin, ampicillin, erythromycin, gentamycin, imipenem, kanamycin, ofloxcin, streptomycin, penicillin, trimethoprim, trimethoprim/sulfamethoxazole, tobramycin, and vancomycin. Multiple resistance patterns of the isolates were observed in TeMiNor Cip (1.4%), TeMiNor (7.1%), TeMiCip (2.9%), TeMiN (1.4%) and TeMi (44.3%). The results of this study indicate that many L. monocytogenes isolates are resistant to antimicrobial agents including Te and Mi. The possibility that the isolates could increasingly acquire multiple antimicrobial resistant properties cannot be precluded.

• 대한수의학회지
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• 제40권2호
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• pp.325-332
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• 2000

The growth characteristics and the cellular protein patterns of the Lactobacillus plantarum isolated and identified from oat silage were examined in order to confirm whether it will be used practically as probiotics or not. L plantarum was identified by morphological and biochemical tests including of final conforming by API 50CHL kit. The cultivation in MRS broth of the strain under the condition of different temperature, proved that they grew into $2.0{\times}10^{9}$ in $25^{\circ}C$, into $1.4{\times}10^{9}$ in $35^{\circ}C$ but they decreased into $4.5{\times}10^{5}$ growth in $45^{\circ}C$. The comparison of the growth by measurement of O.D600nm value after 24 hour cultivation between L plantarum and commercial probiotics, showed that the strain had a higher growth than commercial as 1.841 : 1.623. The measurement of it under bile acid's existence, indicated that this isolation was not influenced by bile acid and the tolerance was $3.2{\times}10^{9}$, $3.9{\times}10^{9}$ and $3.2{\times}10^{9}$, respectively, when each of 0%, 1%, and 2% oxigall existed. The examination of their antibiotics susceptibility by disk diffusion test, proved that L plantarum showed resistance against danofloxacin(5mcg), gentamycin(10mcg), kanamycin(30mcg), neomycin(30mcg) and streptomycin(10mcg). Based upon the test of the bacteriocin formation of this L plantarum, it was found out that the inhibition zone was not formed. In growth of L plantarum and E coli in nutrient broth, all E coli died out within 6 hours after cultures.

• Archives of Pharmacal Research
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• 제24권4호
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• pp.307-311
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• 2001

The activation of NF-$\kappa$B induced by kojic Acid, an inhibitor of tyrosinase for biosynthesis of melanin in melanocytes, was investigated in human transfectant HaCaT and SCC-13 cells. These two keratinocyte cell lines transfected with pNF-$\kappa$B-SEAP-NPT plasmid were used to determine the activation of NF-$\kappa$B. Transfectant cells release the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the NF-$\kappa$B activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selective marker of geneticin resistance. NF-$\kappa$B activation was measured in the SEAP reporter gene assay using a fluorescence detection method. Kojic Acid showed the inhibition of cellular NF-$\kappa$B activity in both human keratinocyte transfectants. It could also downregulate the ultraviolet ray (UVR)-induced activation of NF-$\kappa$B expression in transfectant HaCaT cells. Moreover, the inhibitory activity of kojic Acid in transfectant HaCaT cells was found to be more potent than known antioxidants, e.g., vitamin C and N~acetyl-L-cysteine. These results indicate that kojic Acid is a potential inhibitor of NF-$\kappa$B activation in human keratinocytes, and suggest the hypothesis that NF-$\kappa$B activation may be involved in kojic Acid induced anti-melanogenic effect.

• Laboraroty Animal Research
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• 제34권4호
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• pp.279-287
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• 2018

Placenta specific 8 (PLAC8, also known as ONZIN) is a multi-functional protein that is highly expressed in the intestine, lung, spleen, and innate immune cells, and is involved in various diseases, including cancers, obesity, and innate immune deficiency. Here, we generated a Plac8 knockout mouse using the CRISPR/Cas9 system. The Cas9 mRNA and two single guide RNAs targeting a region near the translation start codon at Plac8 exon 2 were microinjected into mouse zygotes. This successfully eliminated the conventional translation start site, as confirmed by Sanger sequencing and PCR genotyping analysis. Unlike the previous Plac8 deficient models displaying increased adipose tissue and body weights, our male Plac8 knockout mice showed rather lower body weight than sex-matched littermate controls, though the only difference between these two mouse models is genetic context. Differently from the previously constructed embryonic stem cell-derived Plac8 knockout mouse that contains a neomycin resistance cassette, this knockout mouse model is free from a negative selection marker or other external insertions, which will be useful in future studies aimed at elucidating the multi-functional and physiological roles of PLAC8 in various diseases, without interference from exogenous foreign DNA.

• 대한수의학회지
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• 제44권2호
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• pp.217-224
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• 2004

This study was carried out to investigate the antibiotic resistance pattern of Listeria spp. and Staphylococcus aureus. A total of 17 (14.8%) L. monocytogenes, 13 (11.3%) L. innocua, 7 (7%) L. welshimeri, and 83 (72.2%) S. aureus were isolated from commercial poultry carcasses in Seoul and Kyonggi province during the period between 2001 and 2003. Antibiotic susceptibility test of all Listeria strains isolated was performed by the disk agar diffusion method. Antibiotics used in the study were as follows; Amikacin (An), Ampicillin (Am), Cephalothin (Cf), Chloramphenicol (C), Ciprofloxacin (Cip), Erythromycin (E), Gentamicin (Gm), Imipenem (Ipm), Kanamycin (K), Minocycline (Mi), Neomycin (N), Norfloxacin (Nor), Ofloxacin (Ofx), Penicillin (P), Streptomycin (S), Tetracycline (Te), Tobramycin (Nn), Trimethoprim (Tmp), Trimethoprim/Sulfamethoxazloe (Sxt), and Vancomycin (Va). The antibiotic resistance pattern of S. aureus isolates was performed by the disk agar diffusion method. For the latter program, antibiotics used to the study were as follows; Cf, C, Cip, Clindamycin (Cc), E, Gm, Ipm, Nafcillin (Nf), Oxacillin (Ox), P, Te, Sxt, and Va. Of the 17 L. monocytogenes isolates, 94.1% were resistant to Te, 88.2% to Mi, 11.8% to Nor, 11.8% to S, 5.9% to Cip, and 5.9% to C. Of 13 L. innocua, 53.8% were resistant to Te, 23.1% to Mi, 23.1% to S, 7.7% to Cip, and 7.7% to Nor. Of 7 L. welshimeri, 57.1% were resistant to Te, and 14.3% to Am. Of 83 S. aureus, 100% were resistant to Te, 86.7% to Gm, 34.9% to P, 15.7% to Cip, 12% to Cc, 9.6% to E. The multiple antibiotic resistance patterns of L. monocytogenes isolates were observed in Te Mi Cip (5.9%), Te Mi Nor (5.9%), Te Mi (76.5%), and Te Nor (5.9%). Multiple antibiotic resistance was also found in L. innocua isolates. Resistant to Te Mi S Cip Nor was 7.7%, Te Mi S (7.7%), Te Mi (7.7%), and was 7.7% to Te S. Antibiotic resistance patterns for S. aureus isolats were demonstrated to Te Gm P Cip Cc E (6.0%), Te Gm Cip Cc E (3.6%), Te Gm P Cc (1.2%), Te Gm P (15.6%), Te Gm Cip (2.4%), Te P Cip (2.4%), Te Gm Cc (1.2%), Te Gm (56.6%), Te P (9.6%), and to Te Cip (1.2%). The results of this study suggest a high incidence of Lsteria spp. and S. aureus on poultry carcasses. The contaminated poultry carcasses may be a potential vehicle for foodborne infections due to multiple antimicrobial resistant organisms.

• Applied Biological Chemistry
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• 제35권3호
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• pp.179-185
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• 1992

우리나라 남부지방(南部地方)의 5개(個) 산지(山地) 목초지(牧草地)에 분포한 알팔파근류균(根瘤菌) R. meliloti의 밀도(密度)와 실소고정력(室素固定力), 그리고 항생제(抗生劑) 반응(反應) 특성(特性)을 조사하였다. 그 결과(結果), 초지토양(草地土壤)에 분포한 R. meliloti 밀도(密度)는 $1.7{\times}10^2\;cells/g$.토양(土壤)(여주시(麗州市) 충효동(忠孝洞)) 내지 $1.0{\times}10^5\;cells/g$.토양(土壤)(경주군(慶州郡) 감포면(甘浦面)) 수준(水準) 이었으며, 토양 pH와는 정(正)의 상관(相關)을 보였다. 그리고, 초지토양(草地土壤)의 알팔파 접종효과(接種效果)로 본 토착(土着) R. meliloti의 질소고정력(窒素固定力)은 전반적으로 표준균주(標準菌株)인 TAL 접종제(接種劑) 보다 저조(低調)하였으나, 달성군(達城郡) 옥포면(玉浦面) 초지(草地)에서 분리(分離)된 YCKa 539, YCKa 542 2균주(菌株)는 비슷하거나 우수(優秀)한 질소고정력(窒素固定力)을 보였다. 초지토양별로 보면 경주군(慶州郡) 견곡면(見谷面)의 것이 가장 높았고, 다음으로 달성군(達城郡) 옥포(玉浦)>경주시(慶州市) 충효(忠孝)>달성군(達城郡) 화국(花國)>경주군(慶州郡) 감포(甘浦) 균주(菌株) 순(順)이었다. 분리균주(分離菌株)들은 항생제(抗生劑) Ery($100\;{\mu}g/ml$), Nal($160\;{\mu}g/ml$), Str($10\;{\mu}g/ml$)에 대해 각각(各各) 67%, 77%, 80%정도의 내성경향(耐性傾向)을 보이면서 전체적으로는 14개(個)의 항생제(抗生劑) 내성류형(耐性類型)으로 분류(分類)되었는데, 이들 류형(類型)과 균주(菌株)의 질소고정력간(窒素固定力間)에는 뚜렷한 관계경상(關係傾商)을 보이지 않았다. 그리고, 분리균주(分離菌株) 중(中)에서 질소고정력(窒素固定力)이 가장 우수(優秀)하였던 YCKa 542균주(菌株)는 Ery Nal, Neo, Str 내성인 X번형(番型)에 속하였다.