• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.86-86
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• 2003

Research has been in progress for more than a decade to production of useful proteins by genetic modification in cattle. However, the levels of protein production in transgenic cattle have been reported very low. To enhance protein production in transgenic animal, we tried homologous recombination to donor cells for production of transgenic clone cattle through nuclear transfer procedure. Thus, we constructed the two targeting vectors of human thrombopoietin (TPO) at bovine $\beta$-casein locus using homologous recombination with 13.6 kb and 9.6 kb homology. In two targeting vectors, positive selection was through the neomycin resistance gene and negative selection was by the diphtheria toxin (DT). Gene targeting was attempted in bovine embryonic fibroblasts (bEF) and bovine ear skin fibroblasts (bESF). To determine the most appropriate concentration of neomycin for bEF and bESF, G4l8 resistance was confirmed by culturing the cells in various concentrations of the drug and both of the cells were optimally selected at $900 \mu g/ml$ of neomycin. The transfected bEF and bESF by the targeting vectors were colonized efficiently at the ratio of DNA to transfection reagent such as $4 \mu g$:2 ${mu}ell$ and $1 \mu g$:$2 \mu l$. Comparing number of healthy clones from passage 4 to passage 8, bESF (17%) persist in culture for much longer than bEF (6%). The two gene-targeted bESF clones of 30 random-integrated clones with 9.6 kb homology length were confirmed, however, nothing was out of 72 random integration clones with 13.6 kb homology length, The DT also worked more efficiently in clones transfected with the vector of 9.6 kb homology length. Our data suggests that the choice of donor cell for long culture period should be considered to obtain targeted cell clone, and the gene-targeting frequency and the DT working efficiency are dependent on the length of target homology.

• 한국미생물·생명공학회지
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• 제37권3호
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• pp.258-265
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• 2009

축산환경개선용 미생물 제품 중 시장점유율을 기준으로 상위제품 A, B,하위 제품 E, F와 현재 개발완료 되어진 C, D를 선정하여 생균수 측정과, 사료첨가용으로 사용이 금지된 항생제에 대한 내성 테스트를 실시하였다. 생균수 측정 결과 상위 점유율의 A, B사 제품은 표기상의 생균수와 일치하였으나, 하위 점유율을 차지하고 있는 E제품의 경우 함유균주나 효능에 대한 정보를 제공하고 않고 있을뿐더러 F사의 제품은 균수가 적게 측정되어 실제 표기사항과 일치하지 않았다. 항생제 내성실험결과 점유율에 따른 상관관계는 없었으나, B사를 제외한 제품의 대부분에서 내성균이 존재함을 확인하였다. Lincosimides 계열의 Lincomycicne과 Clindamycin의 경우 B사를 제외한 제품에서 내성균이 존재하였다. Penicillins 계열의 Amoxicillin, Ampicillin, Penicillin 및 Macrolide 계열의 Erythromycin 항생제는 B사와 E를 제외한 제품에서 내성균이 존재함을 확인하였다. Quinoline계열 Norploxacin 또한 B사와 E사를 제외한 제품에서 내성균을 보였으며 Neomycin의 항생제 또한 유사한 내성분포 결과를 나타내었다. Neomycin과 같은 Aminiglycosides계열의 Gentamycin, Streptomycin은 B사제품을 제외한 제품에서 내성을 나타내었다. 마지막으로 내성-균으로 사용 금지된 Tetracycline계열의 Oxytetracyclin은 12가지 항생제중 가장 높은 비율의 감수성을 나타내었으나 B, E사를 제외한 제품에서 내성균의 존재를 확인하였다. 실험결과 사용 금지된 항생제 품목임에도 불구하고 많은 제품에서 때 성균이 존재하였으며, 이에 따라 환경개선제 특성의 정확한 표기와 체계적인 유통 체계와 검증 과정이 필요하다고 사료된다.

• 한국식품과학회지
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• 제42권6호
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• pp.643-647
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• 2010

기준 설정 전인 2006년 식품의약품안전청에서 실시한 연구과제의 결과보고서에 의하면 200건 벌꿀 중 네오마이신은 검출되지 않았고, 스트렙토마이신의 경우 23건이 검출되었으며, 아미트라즈(그 대사산물 포함), 코마포스의 경우 각각 10건, 9건의 검출되었다(8). 그러나 기준 설정 후인 본 연구의 모니터링 결과 해당 동물용의약품의 검출율이 현저하게 떨어진 것으로 미루어 볼때 07년 기준 설정 후 설정된 잔류허용기준에 맞춰 관리되고 있음을 알 수 있었다.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-1
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• pp.93-93
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• 2003

RNA plays an important role in numerous biological processes but little is known about the interactions between small organic molecules and RNA. Our previous work has shown that the heterodimeric compound designed by conjugation with neomycin and loop-specific chemical bind to the stem-loop structured RNA motifs. In the present study, heterodimer was used, in a reverse way, as a probe to identify structured RNA motifs. (omitted)

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.161.2-161.2
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• 2003

In order to investigate the pathogenic genes and genetic relationships of Y. pseudotuberculosis, we isolated 9 strains of Y. pseudotuberculosis from about 380 spring water sites in Seoul and carried out antibiotic susceptibility test, biological test and molecular typing. All isolated strains were distributed throughout the northeast area in Seoul (Mt. Bookhan, Mt. Soorak, Mt. Boolam and etc...).Antibiotic susceptibility test revealed that all the strains were susceptible to chloramphenicol, gentamicin, neomycin and amoxicillin/clavulanic acid, but were resistant to novobiocin and vancomycin. (omitted)

• 한국식품과학회지
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• 제31권5호
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• pp.1357-1362
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• 1999

한국인의 장내세균중의 유해효소생성 균주를 분리하여 장내환경 개선연구에 활용하고자 ${\beta}-glucuronidase$와 $7{\alpha}-dehydroxylase$의 유해효소를 분비하는 장내세균으로 알려진 Clostridium속 균주를 분리하였다. 동일인의 장내세균을 2회에 걸쳐서 검색한 후 최다 우점종이외의 장내세균은 동일인이라도 식이등 외부환경에 의하여 변화될 수 있음을 알았다. 이어서 Neomycin-Nagler 선택배지를 사용하여 Clostridium spp.를 14종 분리하여 이들 균주가 유해효소를 생산하는지를 조사하였다. 일차 선발균주중 Clostridium sp. Isolate-11은 ${\beta}-glucuronidase$를 강력히 생성하여 0.021 unit/mg protein의 활성을 보여주었으나 이들 분리 균주중에서 $7{\alpha}-dehydroxylase$의 역가를 TLC상에서 검출할 수 있는 균주는 발견하지 못했다. Isolate-11를 배양생리학적으로 동정한 결과 이 균은 Clostridium scatologenes인 것으로 잠정동정하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권9호
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• pp.1336-1342
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• 2005

The present research work was conducted to evaluate the beneficial effects as well as the safety aspects of lactobacilli as probiotic. Lactobacilli were isolated from poultry faecal samples, feed samples and from some known preparations procured from poultry feed manufacturers. L. acidophilus and L. sporogenes were tested for the antibacterial activity against four poultry pathogens viz. Escherichia coli, Salmonella spp., Proteus spp. and Pseudomonas aeruginosa. Cell free supernatant (CFS) of L. acidophilus exhibited significantly higher antibacterial activity against Salmonella spp. at original pH (4.50${\pm}$0.02). At the adjusted pH (6.50${\pm}$0.02) significantly higher antibacterial activity was recorded against indicator organism except for P. aeruginosa. Likewise, L. sporogenes exhibited similar antibacterial activity at original as well as adjusted pH except for E. coli. Antibacterial activity against E. coli was significantly higher at adjusted pH than at original pH of CFS. The competitive exclusion of E. coli by lactobacilli over the intestinal epithelial cells (IEC) was checked. L. acidophilus strain I, which was of poultry origin, exhibited maximum attachment over IEC as compared to other three strains of non-poultry origin viz. L. acidophilus strain II, L. sporogenes strain I and II. Overall, L. acidophilus exhibited higher competitive exclusion as compared to L. sporogenes. All the lactobacilli of poultry origin were most sensitive to penicillin G, amoxycillin, ampicillin and chloramphenicol, least sensitive to sulphamethizole, ciprofloxacin, neomycin, norfloxacin and pefloxacin and resistant to metronidazole and nalidixic acid. The isolates from probiotic preparations were most sensitive to ampicillin, amoxycillin and tetracycline, least sensitive to sulphamethizole, norfloxacin, neomycin and ceftriazone and resistant to nalidixic acid and metronidazole. Eight of the multiple drug resistant lactobacilli isolates were studied for the presence of plasmids. Plasmids could be extracted from six isolates of lactobacilli. These plasmids could be responsible for bacteriocin production or for antibiotic resistance of the strains. The lactobacilli need further studies regarding their safety for use in the probiotic preparations.

• 한국동물위생학회지
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• 제23권1호
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• pp.61-69
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• 2000

Thirty-one dairy cow from two farm(more than 500,000 cells/ml of bulk milk) in Kyongbuk northern province were selected because of their high somatic cell(more than 500,000 cells/ml of milk In individual cow). Each cow received. staphylococcus aureus vaccine(Labac Staph) and immunostimulant(Ultracon) by intramuscular injection to be repeated every fifteen days for S times. The present study was investigated variation of somatic cell after administration of Labac Staph and Ultracon, and antibiotics resistance of isolated staphylococcus spp from milk in selected cow. The results obtained through the survey were summarized as follows ; 1. Ten dairy cow was injected in A farm. Chronic mastitic two cow after 2rd injection was weeded out the herd. Decrease rate of somatic cell after 1st, 2nd, ,3rd, 4th and 5th administration were 41.4%, 35.6%, 56.4%, 65.4% and 36.7%, respectively. Twenty-one cow was injected in B farm. Chronic mastitic five cow after ,3rd injection was weeded out the herd. Decrease rate of somatic cell after 1st, 2nd, 3rd, 4th and 5th administration were 36.9%, 59.9%, 24.5%, 62.6% and 78.4%, respectively. 2. In A farm, isolated staphylococcus spp were identified as S hyicus 2 strains(11.8%), coagulase negative staphylococcus 15 stains(89.2%) and S epidermidis 6strain(35.3%). In B farm, isolated staphylococcus spp were identified as S aureus 19 strains(55.98%) and coagulase negative staphylococcus 15 strains (44.2%). 3. In A fm, antibiotics resistant rate of isolated staphylococcus spp was high at ampicillin, penicillin and kanamycin, and middle at neomycin, streptomycin and erythromycin. in B farm, antibiotics resistant rate was moderate at ampicillin, penicillin, gentamicin, ka-namycin, neomycin, streptomycin, erythromycin and tetracycline, and coagulase negative staphylococcus spp was moderate at streptomycin.

• 식물조직배양학회지
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• 제28권1호
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• pp.53-58
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• 2001

제초제 저항성 오이 (Cucumis sativus L. cv Green angel)를 생산하기 위하여 배발생 현탁배양세포와 binary vector pGA-bar을 지닌 Agrobacterium tumefacians (LBA4404)를 공동배양하였다. 형질전환 벡터의 T-DNA부분에는 kanamycin에 저항성을 나타내는 neomycin phosphotrans ferase (npt II) 유전자와 phosphinothricin (PPT)에 저항성을 나타내는 phosphinothricin acetyltransferase (bar) 유전자를 지니고 있다. 48시간의 공동배양 후 배발생 캘러스는 20mg/L PPT가 함유된 성숙배지에서 배양하였다. 약 200개체의 형질전환 유식물체를 40mg/L PPT가 첨가된 호르몬이 없는 배지에서 생산하였다. 5개의 오이 형질전환 식물체의 염색체에 bar유전자가 도입되어 발현되는 것을 northern blot 분석을 통하여 확인하였다. 형질전환 오이 식물체가 토양에서 성숙되었다. 성숙한 오이 식물체는 PPT가 함유된 상업적 제초제 (Basta)를 일반적인 사용 농도 (3ml/L)처리시에도 저항성을 나타내며 생장하였다.

• 대한화장품학회지
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• 제27권1호
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• pp.17-27
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• 2001

A cell-based assay system for monitoring NF-$textsc{k}$B activity was developed to determine the influence of activated NF-$textsc{k}$B in human HaCaT cells. The pNF-$textsc{k}$B-SEAP-NPT plasmid that permits expression of the secreted alkaline phosphatase (SEAP) reported gene in response to the NF-$textsc{k}$B activity and contains neomycin phosphotransferase (NPT) gene for the geneticin resistance in host cells was constructed and transfected into human keratinocyte cell line HaCaT. Human HaCaT transfectant cells secreted the SEAP enzyme into the culture medium in a time-dependent manner until 72h. NF-$textsc{k}$B activities were measured in the SEAP reporter gene assay using a fluorescent detection method. The treatment of HaCaT cell transfectants with known antioxidants [e.g., N-acetyl-L-cysteine and vitamin C] showed inhibition of NF-$textsc{k}$B activity in a time-and concentration-dependent manner. The phorbol 12-myristate 13-acetate (PMA) known as a stimulator of NF-$textsc{k}$B expression demonstrated that it increased NF-$textsc{k}$B activity in a time- and concentration-dependent manner. This assay system could be used to determine the quantitative measurement of NF-$textsc{k}$B activity in the human skin and allow the screening of anti-inflammatory agents from various synthetic chemicals and natural products for dermatological purpose. Abbrevitions used: NF-$textsc{k}$B, nuclear factor kappa B; I-$textsc{k}$B, Inhibitory kappa B; SEAP, secreted alkaline phosphatase; NPT, neomycin phosphotransferease; PCR, polymerase chain reaction: dNTP, deoxynucleoside triphosphates; DMEM, dulbecco’s modified eagle medium; FBS, fetal bovine serum; PBs, phosphate-buffered saline; MUP, 4-methylumbellifery phosphate; NAC, N-acetyl-L-cysteine; DMSO, dimethyl sulfoxide; PMA, phorbol 12-myristate 13-acetate.