• Title/Summary/Keyword: STR

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Evaluation of Antibiotics Resistance for Human-harmful Bacteria Isolated from Eco-friendly and Practical Cultivation Farms of Hot Pepper and Tomato (고추 및 토마토 친환경 및 관행재배지에서 분리한 인체 유해세균의 항생제 저항성 평가)

  • Lee, Sung-Hee;Do, Jiwon;Kim, Seong Kyeom;Oh, Kwang Kyo;Park, Jae-Ho
    • Korean Journal of Organic Agriculture
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    • v.31 no.4
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    • pp.381-394
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    • 2023
  • This study was conducted to monitor the antibiotics resistance of human-harmful bacteria isolated in the agricultural environment for hot peppers (Capsicum annuum) and tomato (Lycopersicon esculentum). As a result, we isolated 120 bacterial species (34 on fruits, 48 in soil, 21 in water, and 17 in manure), identified them with the 16S rRNA sequence, analyzed minimum inhibitory concentration (MIC) for 26 antibiotics using Sensititre ARIS Hi-Q system and then evaluated whether each bacterial genus acquired resistance for the tested antibiotics or not, according to the CLSI criteria. From difference in MIC between eco-friendly (EFM) and practical (PFM) cultivation farms, Klebsiella spp. isolated from EFM was resistant to ampicillin (AMP) and nalidixic acid (NAL), and that isolated from PFM was resistant to streptomycin (STR) and tetracycline (TET). Enterobacter spp. isolated from EFM was resistant to AMP and azithromycin (AZI), and that isolated from PFM was resistant to AMP, AZI, and STR. Meanwhile, Pseudomonas spp. isolated from EFM and PFM were all resistant to AMP, AZI, cefotaxime (FOT), cefoxitin (FOX), ceftriaxone (AXO), CHL, NAL, and STR. Staphylococcus spp. isolated from EFM and PFM were resistant to gentamycin (GEN), STR, and kanamycin (KAN), and in particular, that from EFM showed resistance for erythromycin (ERY). In conclusion, our study suggested that EFM lead STR antibiotics resistance for human-harmful bacteria to decrease, because only the bacteria isolated from hot pepper and tomato crop with PFM have showed resistance against STR antibiotics, regardless of bacterial genus.

Validation of QF-PCR for Rapid Prenatal Diagnosis of Common Chromosomal Aneuploidies in Korea

  • Han, Sung-Hee;Ryu, Jae-Song;An, Jeong-Wook;Park, Ok-Kyoung;Yoon, Hye-Ryoung;Yang, Young-Ho;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • v.7 no.1
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    • pp.59-66
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    • 2010
  • Purpose: Quantitative fluorescent polymerase chain reaction (QF-PCR) allows for the rapid prenatal diagnosis of common aneuploidies. The main advantages of this assay are its low cost, speed, and automation, allowing for large-scale application. However, despite these advantages, it is not a routine method for prenatal aneuploidy screening in Korea. Our objective in the present study was to validate the performance of QF-PCR using short tandem repeat (STR) markers in a Korean population as a means for rapid prenatal diagnosis. Material and Methods: A QF-PCR assay using an Elucigene kit (Gen-Probe, Abingdon, UK), containing 20 STR markers located on chromosomes 13, 18, 21, X and Y, was performed on 847 amniotic fluid (AF) samples for prenatal aneuploidy screening referred for prenatal aneuploidy screening from 2007 to 2009. The results were then compared to those obtained using conventional cytogenetic analysis. To evaluate the informativity of STR markers, the heterozygosity index of each marker was determined in all the samples. Results: Three autosomes (13, 18, and 21) and X and Y chromosome aneuploidies were detected in 19 cases (2.2%, 19/847) after QF-PCR analysis of the 847 AF samples. Their results are identical to those of conventional cytogenetic analysis, with 100% positive predictive value. However, after cytogenetic analysis, 7 cases (0.8%, 7/847) were found to have 5 balanced and 2 unbalanced chromosomal abnormalities that were not detected by QF-PCR. The STR markers had a slightly low heterozygosity index (average: 0.76) compared to those reported in Caucasians (average: 0.80). Submicroscopic duplication of D13S634 marker, which might be a unique finding in Koreans, was detected in 1.4% (12/847) of the samples in the present study. Conclusion: A QF-PCR assay for prenatal aneuploidy screening was validated in our institution and proved to be efficient and reliable. However, we suggest that each laboratory must perform an independent validation test for each STR marker in order to develop interpretation guidelines of the results and must integrate QF-PCR into the routine cytogenetic laboratory workflow.

The development of anti-DR4 single-chain Fv (ScFv) antibody fused to Streptavidin (Streptavidin이 융합된 DR4 항원에 특이적인 single-chain Fv 항체의 개발)

  • Kim, Seo Woo;Wu, Sangwook;Kim, Jin-Kyoo
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.330-342
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    • 2018
  • The Streptavidin and Biotin system has been studied most extensively as the high affinity non-covalent binding of Biotin to STR ($K_D=10^{-14}M$) and four Biotin binding sites in tetrameric Streptavidin makes this system useful for the production of multivalent antibody. For the application of this system, we cloned Streptavidin amplified from Streptomyces avidinii chromosome by PCR and fused to gene of hAY4 single-chain Fv antibody specific to death receptor 4 (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand. The hAY4 single-chain Fv antibody fused to Streptavidin expressed in Escherichia coli showed 43 kDa monomer in heated SDS-PAGE. However, this fusion protein shown in both non-heated SDS-PAGE and Size-exclusion chromatography exhibited 172 kDa as a tetramer suggesting that natural tetramerization of Streptavidin by non-covalent association induced hAY4 single-chain Fv tetramerization. This fusion protein retained a Biotin binding activity similar to natural Streptavidin as shown in Ouchterlony assay and ELISA. Death receptor 4 antigen binding activity of purified hAY4 single-chain Fv fused to Streptavidin was also confirmed by ELISA and Westernblot. In addition, surface plasmon resonance analysis showed 60-fold higher antigen binding affinity of the hAY4-STR than monomeric hAY4 ScFv due to tetramerization. In summary, hAY4 single-chain Fv fused to Streptavidin fusion protein was successfully expressed and purified as a soluble tetramer in E. coli and showed both Biotin and DR4 antigen binding activity suggesting possible production of bifunctional and tetrameric ScFv antibody.

Performance of MiniPCRTM mini8, a portable thermal cycler (휴대용 DNA증폭기 MiniPCRTM mini8 Thermal Cycler의 성능 검토)

  • Kwon, Han-Sol;Park, Hyun-Chul;Lee, Kyungmyung;An, Sanghyun;Oh, Yu-Li;Ahn, Eu-Ree;Jung, Ju Yeon;Lim, Si-Keun
    • Analytical Science and Technology
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    • v.29 no.2
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    • pp.79-84
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    • 2016
  • A small and inexpensive thermal cycler (PCR machine), known as the MiniPCRTM Mini8 Thermal Cycler (Amplyus, Cambridge, MA, USA), was developed. In this study, the performance of this PCR machine was compared with the GeneAmp® PCR system 9700 (Applied Biosystems) using four autosomal short tandem repeat (STR) kits, a Y-chromosome STR kit, and a mitochondrial DNA HV1/HV2 sequence analysis. The sensitivity and stochastic effects of the STR multiplex kits and the quality of the DNA sequence analysis were similar between the two PCR machines. The MiniPCRTM Mini8 Thermal Cycler could be used for analyses at forensic DNA laboratories and crime scenes. The cost of the PCR is so economical that school laboratories and individuals could use the machines.

Effect of peptide on the mixed fermentation of Lactobacillus helveticus YM-1 and Streptococcus lactis $ML_3$ in skim milk (Lactobacillus helveticus YM-1과 Streptococcus lactis $ML_3$의 혼합발효에 미치는 peptide의 영향)

  • 박정길;류인덕;유주현
    • Microbiology and Biotechnology Letters
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    • v.14 no.6
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    • pp.487-493
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    • 1986
  • Lactobacillus helveticus YM-1and Streptococcus lactis Ml$_3$ were inoculated together in reconstituted non-fat skim milk medium, and then their proteolytic activity and stimulatory compound for acid production were investigated. Significant difference between Lactobacillus helveticus YM-1 and Streptococcus lactis Ml$_3$was observed in the proteolytic activities. The proteolytic activity of Lactobacillus helveticus YM-1 and Streptococcus lactis Ml$_3$ was 105 $\mu\textrm{g}$/$m{\ell}$ and 30 $\mu\textrm{g}$/$m{\ell}$ when converted the amounts of hydrolysates of milk protein determined by Folin Ciocaleau phenol method into their tyrosine equivalent Stimulatory compounds in cell-free filtrate of Lactobacillus helveticus YM-1were identified as peptide with a molecular weight of approximately 4, 300 for the acid production by Streptococcus lactis Ml$_3$. Some kinds of amino acids, such as histidine, lysine, arginine and glutamic acid, were rich in acid hydrolysates of peptide. Among amino acids, histidine, glutamic acid and phenylalanine stimulated acid production, on the contrary isoleucine inhibited.

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Allele Frequency of the Short Tandem Repeat(STR) Loci FFv Triplex(F13A01, FESFPS, vWA) Gene by Polymerase Chain Reaction in the Korean Population (한국인에서 중합효소 연쇄반응법에 의한 Short Tandem Repeat(STR) 유전좌위 FFv Triplex(F13A01, FESFPS, vWA) 유전자빈도 검색)

  • Yoon, Chang-Lyuk;Ryu, Geun-Chun
    • Journal of Oral Medicine and Pain
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    • v.24 no.3
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    • pp.335-345
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    • 1999
  • 법의학적 개인식별 및 친생자 감정시 여러 개의 single tandem repeats(STR) 유전좌위 검색이 필요하다. 그 이유는 STR 유전좌위는 대립유전자 수가 적고 이형접합도가 낮아 서로 다른 개체간에도 동일한 유전좌위를 가질 확률이 높기 때문에 개인식별에 대한 기여도가 떨어지게 된다. 따라서 여러 개의 다양한 STR 유전좌위들을 동시에 분석함으로써 우연적으로 개체간에 유전자형이 일치할 가능성을 낮추어야 감정의 신뢰성을 높일 수 있으며 이에는 각 STR 유전좌위에 대한 유전좌위의 분포가 인종별, 지역별로 달라 이에 대한 유전자분포를 구하는 것이 선행조건이다. 이에 본 연구에서는 법의학적 개인식별 및 친자감정시 기초자료로 활용하기 위하여 서로 혈연관계가 없는 201명의 한국인 혈액에서 DNA를 추출하여 STR 유전좌위증 human coagulation factor XIII A subunit gene(F13A0l Locus), human c-fes/fps proto-oncogene(FESFPS Locus), human von Willebrand factor gene (vWA Locus)등 FFv Triplex 유전자를 중합효소반응에 의하여 동시에 증폭하고, 폴리아크릴아마이드겔을 이용한 전기영동 및 질산은 염색을 시행한 후 FFv Triplex유전자의 유전자형 및 대립유전자 빈도 등을 분석하여 다음과 같은 결과를 얻었다. (1) F13A01유전자는 5개의 대립유전자, 12개의 유전자형을 검출하였으며, 이형접합도는 60.7%로 나타났고 대립유전자 및 유전자빈도는 3.2, 4, 5, 6, 16 대립유전자에서 각각 0.34 3, 0.114, 0.062, 0.475, 0.005로 나타났으며, 대립유전자 7, 8, 9, 10, 11, 12, 13, 14, 15는 검출되지 않았다. (2) F13A01 대립유전자다양성 (allelic diversity value)은 0.641, 개인식별력(PD)은 0.814를 보였으며 대립유전자다양성 및 이형접합도가 다른 민족과 비교할 때 다소 낮았다. (3) FESFPS유전자는 8개 대립유전자 모두 나타났으며, 15개의 유전자형을 검출하였으며, 이형접합도는 66.7%로 나타났고 대립유전자 및 유전자빈도는 7, 8, 9, 10, 11, 11, 12, 13, 14 대립유전자에서 각각 0.002, 0.002, 0.005, 0.032, 0.507, 0.264, 0.197, 0.007로 나나났다. (4) FESFPS 대립유전자다양성(allelic diversity value)은 0.641, 개인식별력(PD)은 0.804를 보였다. (5) vWA유전자는 9개의 대립유전자, 23개의 유전자형을 검출하였으며, 이형접합도는 80.1%로 나타났고 대립유전자 및 유전자 빈도는 11, 12, 14, 15, 16, 17, 18, 19, 20 대립유전자 에서 각각 0.002, 0.002, 0.219, 0.032, 0.187, 0.279, 0.189, 0.072, 0.017로 나타났으며, 대립 유전자 13, 21는 검출되지 않았다. (6) vWA 대립유전자다양성(allelic diversity value)은 0.799, 개인식별력(PD)은 0.924로 매우 높게 나타냈다.

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Effect of Chlorella Extract on Acid Production and Growth of Yoghurt Starter (Chlorella 추출물 첨가가 Yoghurt Starter의 산 생성 및 증식에 미치는 영향)

  • 조은정;남은숙;박신인
    • The Korean Journal of Food And Nutrition
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    • v.17 no.1
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    • pp.8-17
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    • 2004
  • The effect of chlorella extract on the growth and acid production of yoghurt starter was investigated in order to prepare the yoghurt added with chlorella extract. The various levels of chlorella extract powder were added to skim milk medium and the medium was fermented by single or mixed culture of 4 types of lactic acid bacteria such as Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus bulgaricus. The changes in acid production(pH, titratable acidity) and number of viable cells of the medium during fermentation in skim milk added with chlorella extract powder have determined. When chlorella extract powder was added to skim milk medium at the levels of 0.5%, 1.0%, 2.0%, and 3.0%, the addition of 0.5% chlorella extract powder with the single culture of Str. thermophilus, Lac. casei, and Lac. bulgaricus showed the highest number of viable cell counts after 9 hours incubation. And also all single cultures of the yoghurt starter produced the higher amounts of acid with the addition of 0.5% chlorella extract powder. When chlorella extract powder was added to the medium at the levels of 0.25%, 0.5%, 1.0%, and 2.0%, the addition of lower lever(0.25∼0.5%) of chlorella extract powder with the mixed culture of the lactic acid bacteria showed more the acidity of pH and the number of viable cell counts. Among the treatments tested, the addition of 0.25% chlorella extract powder with the mixed culture of Str. thermophilus and Lac. casei produced the highest number of viable cell counts after 12 hours incubation. Therefore it was suggested to manufacture the yoghurt with the addition of 0.25% chlorella extract powder and the inoculation of mixed culture of Str. thermophilus and Lac. casei for on the stimulation of growth of the yoghurt starter.

Investigations Into More Exact Weightings of Customer Demands in QFD

  • Crostack, H.A.;Hackenbroich, I.;Refflinghaus, R.;Winter, D.
    • International Journal of Quality Innovation
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    • v.8 no.3
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    • pp.71-80
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    • 2007
  • Apart from the customer demands themselves, the weightings of the customer demands are one of the main input data of a QFD (Quality Function Deployment) and furthermore of the actual construction process of products. Up to now, most interviews with stakeholders have been carried out with questionnaires and then absolute weightings have been used. Now it has been analysed if the use of other interview and evaluation techniques, e.g. relative weightings and Analytic Hierarchy Process (AHP), can improve the precision of the demands and wishes of the stakeholders. Now the task was to analyse if the use of relative weightings as input of a QFD is possible at all, how they have to be adapted and if an increase in precision compared to the use of absolute weightings is reached. When using AHP during the product development it has become clear that only up to seven demands can be rated at the same time by customers. That means that a kind of hierarchy has to be developed to correctly transfer the demands and their weightings into the QFD.

Allele Frequency of the Short Tandem Repeat Locus Human Lipoprotein Lipase(LPL) Gene by Polymerase Chain Reaction in the Korean Population (한국인에서 중합효소연쇄 반응법에 의한 STR 유전좌위 LPL의 유전자빈도 검색)

  • Na, Yun-Ju;Hur, Woong;Yoon, Chang-Lyuk
    • Journal of Oral Medicine and Pain
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    • v.22 no.2
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    • pp.253-260
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    • 1997
  • 한국인 집단에서 개인식별의 기초자료로 활용하고자 한국인 201명을 대상으로 STR 유전좌위 중 하나인 LPL 유전좌위의 유전자 빈도 및 유전자형 분포를 구하였다. 혈액으로부터 추출한 핵 DNA를 중합효소연쇄반응으로 증폭시키고 폴리아크릴아마이드 겔 상에서 전기영동하여 은염색한 후 관찰하여 다음의 결과를 얻었다. 1. 한국인 집단 201명의 LPL 유전자에서 5개의 대립유전자, 7개의 유전자형을 검출하였으며, 이형접합도는 50.7%로 나타났고 대립 유전자다양성 (allelic diversity value)은 0.454, 개 인식 별력 (PD)은 0.674를 보였다. 2. 대립 유전자 및 유전자빈도는 9, 10, 11, 12, 13 대립 유전자에서 각각 0.020, 0.714, 0.100, 0.164, 0.002로 나타났으며, 대립유전자 7, 8, 14는 관찰되지 않았다. 이상의 결과를 볼 때 한국인 집단에서 STR LPL유전좌위의 유전자빈도는 친자감정 등 개인식별에 유용하게 사용할 수 있으나 감정실무에 응용시 다수의 STR유전좌위 및 VNTR유전좌위의 분석을 병행하여야 할 것으로 사료된다.

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