• Title/Summary/Keyword: 계대배양

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Growth Characteristics and Physiological Properties in Milk of Lactobacillus casei CU2604 Isolated from Adult Feces (성인으로부터 분리된 Lactobacillus casei CU2604의 우유배지에서의 생장 특성 및 생리적 특성)

  • Kim, Hee-Jin;Choi, Jae-Kyoung;Lee, Kyung-Min;Im, Jung-Hyun;Eom, Seok-Jin;Kim, Geun-Bae
    • Food Science of Animal Resources
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    • v.29 no.5
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    • pp.619-626
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    • 2009
  • As a trial for the development of a new starter culture for yogurt products, more than two hundred lactic acid bacteria strains were isolated from raw milk and healthy human feces. The strains that showed excellent growth and acid production ability in the 10% skim milk media were selected and identified as Lactobacillus casei through the API carbohydrate fermentation pattern and 16S rDNA sequence analysis. L. casei CU2604 was further investigated for its physiological characteristics as a starter culture compared with a commercial strain. The CU2604 strain showed good acid production and growth characteristics in milk, which were comparable to those of the L. casei Shirota strain. Despite the fact that both these strains displayed the same sugar fermenting pattern and PFGE band pattern, and had similar growth characteristic in milk, L. casei CU2604 exhibited different fatty acid composition in the cell wall, showed more tolerance to bile and to pH, and presented better growth inhibition activity against pathogenic bacteria. Based on these results, the L. casei CU2604 strain holds great promise for use as a novel and efficient starter culture in the production of yogurt. Additional studies on the probiotic characteristics of this strain are currently being conducted.

Differential Potential of Stem Cells Following Their Origin - Subacromial Bursa, Bone Marrow, Umbilical Cord Blood - (줄기세포의 분화능의 기원에 따른 비교 - 견봉하 점액낭, 골수, 탯줄 혈액 -)

  • Sim, Sung Woo;Moon, Young Lae;Kang, Jung Hun
    • Clinics in Shoulder and Elbow
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    • v.15 no.2
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    • pp.65-72
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    • 2012
  • Purpose: To evaluate the differentiation potential of stem cells and their immunophenotype from 3 different sources. Methods: Our study involved three stem cell sources-subacromial bursal tissue, bone marrow, and umbilical cord blood. We obtained the subacromial bursal tissue and bone marrow from the patients undergoing shoulder surgery. After collecting the sample, we applied specific induction media for neurogenic, adipogenic and osteogenic differentiation. Also, flow-cytometry analysis was done to reveal the cell surface antigens. Results: We obtained 100% (8 cases) neural and adipogenic differentiation, but 62.5% (5 of 8 cases) osseous differentiation among the subacromial bursal tissue group. Bone marrow derived cells showed 100% neural (6 cases) and adipogenic (5 cases) differentiation, but 80% (4 of 5 cases) osseous differentiation. Umbilical cord blood derived cells revealed 97% (65 of 67 cases) neural, 53.7% (29 of 54 cases) adipogenic and 68.4% (39 of 57 cases) osseous differentiation. Immunophenotype analysis revealed that surface markers of bone marrow, subacromial bursal cell and umbilical cord blood derived mesenchymal stem cells are different from each other. Conclusions: Mesenchymal stem cells are potential agents in regenerative medicine and are characterized by expression of surface markers and by their differentiation potential. Our study with stem cells from subacromial bursal tissue, bone marrow and umbilical cord discovered that each stem cell has unique differentiation potential and function based on its origin. Various stem cells show multi-lineage differentiations in vitro which can be correlated to in vivo conditions.

The Use of the Pathogen-specific Bacteriophage BCP8-2 to Develop a Rice Straw-derived Bacillus cereus-free Starter Culture (단일 박테리오파지를 이용한 볏짚 유래 Bacillus cereus free 스타터 컬쳐의 개발)

  • Bandara, Nadeeka;Chung, Seo-Jin;Jeong, Do-Youn;Kim, Kwang-Pyo
    • Korean Journal of Food Science and Technology
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    • v.46 no.1
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    • pp.115-120
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    • 2014
  • The purpose of this study was to develop a rice straw-derived Bacillus cereus (B. cereus)-free starter culture for traditional soybean fermented products using a B. cereus-specific bacteriophage, BCP8-2. To determine the optimal medium that supports the growth of rice straw-derived microorganisms and BCP8-2 activity, 5 different culture media were tested. The 5% ground bean (GB) medium was selected for further study. No B. cereus was detected in the BCP8-2-treated rice straw in GB medium, whereas B. cereus at a level of $10^7$ CFU/mL was recovered in the no-phage control. The total bacterial count reached approximately $10^9$ CFU/mL regardless of phage addition. When the 16S rRNA sequence-based microbial community was monitored using denaturing gradient gel electrophoresis (DGGE) and pyrosequencing, a similar microbial community was observed in the phage-treated and control samples. In conclusion, we demonstrate that phage can be used to prepare a rice straw-derived B. cereus-free starter culture with minimal effect on natural microflora.

Effects of established cell lines on bovine embryo development during in vitro culture (계대세포를 이용한 소 수정란의 체외배양 체계에 관한 연구)

  • Joo, Suk-chun;Lee, Byeong-chun;Lee, Won-yu;Choi, Yun-seok;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.647-659
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    • 1997
  • To overcome the difficulties of collecting and culture of primary cell from genital tract on embryonic development, the present study was carried out to investigate the critical effect of cell lines, such as BRL and Vero cell and its conditioned medium on the development of early Korean native cattle embryos in vitro. Oocytes collected from slaughterhouse ovaries were matured in TCM199 containing FSH, estradiol-$17{\beta}$ and FBS with granulosa cell monolayer for 24 hours and then fertilized in vitro using frozen-thawed, heparin-treated spermatozoa in TALP for 30 hours. And then early embryos (1-2cell) were cultured in TCM199 containing 10% FBS with BOEC, Granulosa, BRL, Vera cell monolayers and conditioned medium for 2~3 days. Development to morulae and blastocysts were recorded, also examined the number of blastomeres presented a valuable parameter for the evaluation of embryonic development. The early cleavage rates of in vitro fertilized embryos co-cultured, there was no differences between primary cell and cell lines(p<0.05). The rate of development to the later stage, coculture of BRL cell was significantly higher than that of the primary cell(p<0.05). The rates of development to morula and blastocyst were significantly higher in vero cell than BRL, Granulosa, Oviduct epithelial cell conditioned medium. In the result of effect of serum on development of early bovine embryos, the use of media containing serum were significantly higher than the use of not containing one on development of early and later stage of embryos. The result of number of blastomeres in blastocysts, there is no differences between primary cell and cell lines. The blastocysts from coculture were higher than from conditioned medium in blastomere cells. In summary, these experments have proved that the culture system in TCM199 with BRL, Vero cell monolayers is effective on in vitro development of early bovine embryos, In addition, it is effective to development of bovine embryos that containing serum in conditioned medium, or in co-culture rather than in conditioned medium alone. The use of cell lines opponent to primary cells is effective in bovine embryo culture.

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The Cadmium Biosorption Mechanism in Gram Negative Bacteria, Serratia marcescens (Gram 음성 세균인 Serratia marcescens에 의한 카드뮴 흡착 기작)

  • 이호용;민봉희;최영길
    • The Korean Journal of Ecology
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    • v.22 no.1
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    • pp.39-43
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    • 1999
  • Serratia marcescens, an enterobacterium of gram-negative bacteria, is characterized by resistance of the admium. Cadmium sensitive PM strain did not grow in the medium at cadmium concentration of 50 ppm. PA strain was induced to accommodate to cadmium by cultivating the mother strain (PC strain) in the medium with 50 ppm cadmium. As compared with PC and PM strains, PA strain revealed the excellent growth in cadmium media and accumulated four to five times higher cadmium concentration in cell than other strains. PA strain accumulated 23% of cadmium in cells when cultured in medium treated with 100 ppm cadmium and this cadmium was more accumulated in cytosol fractions than membrane fractions. Analysis by TEM indicated that cadmium was concentrated as a form of granule in cytosol. In protein patterns of cell after the treatment of cadmium, two inducible proteins (28 KDa and 64 KDa) and one reducible protein (45 KDa) were detected by SDS-PAGE. By Atomic Absorption Spectrophotometer, the amounts of cadmium attached to inducible proteins of 28 KDa and 64 KDa were 318.28 ㎍ and 325.37 ㎍ per gram of protein, respectively. It is assumed that these inducible proteins play an important role in the mechanism of cadmium accumulation in cells. A plasmid of 23Kbp was found in S. marcescens. The ability of resistance to cadmium in plasmid was confirmed by curing experiments.

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Nucleotide Sequence Analysis of the Hemagglutinin-Neuraminidase Gene of Urabe AM-9 Strain (Urabe AM-9 볼거리 백신주의 Hemagglutinin-Neuraminidase 유전자 염기서열 분석)

  • Lee, Joo Yeon;Kim, Jee Hee;Lee, Jin Soo;Park, Ji Ho;Sohn, Young Mo
    • Pediatric Infection and Vaccine
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    • v.7 no.1
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    • pp.83-93
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    • 2000
  • Purpose : Urabe AM-9 strain was known to be associated with increased aseptic meningitis. The reason for high incidence of vaccine-associated meningitis was known that nucleotide(nt) substituted form G to A at position 1081 of the hemagglutinin-neuraminidase(HN) gene and therefore, glutamic acid changed to lysine at amino acid 335. We assessed by comparing nt sequence of the HN gene form Urabe AM-9 strain with wild strain and documented the correlation between nt substitution and vaccine-associated meningitis. Methods : Two lots of Urabe AM-9 vaccine distributed in Korea and mumps wild strains isolated from 1998 through 1999 were analysed. Analysis was made by nt sequencing following amplification of HN gene by RT-PCR. Results : Nucleotide substitution at position 343, 1476, 1570 was not found in both Urabe AM-9 vaccines and wild strains. But analysis of vaccine strains and wild strains isolated from patients revealed substitution from G to A at nt 1081 of the HN gene. Therefore, it encodes lysine instead of glutamic acid at amino acid 335. There was no mixture from of G and A at nt 1081. Nt at 1470 of one lot of Urabe AM-9 vaccines changed from C to A after Vero cell passage. Nt at 1727 of vaccines and wild strains was substituted A to G, so it encodes glycine instead of aspartic acid. Conclusion : Nucleotide analysis of HN gene revealed that nt 1081 of Urabe AM-9 vaccines and wild strains had wild type AAA($Lys^{335}$) instead of variant type GAA($Glu^{335}$). The results of this study suggest that there was a probability of vaccine-associated meningitis due to Urabe AM-9 in Korea before. But incidence of actual side effect was not evaluated because there was no reporting system in Korea.

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In Vitro Plant Regeneration for Mass Propagation of Epimedium koreanum Nakai (삼지구엽초의 다량번식을 위한 기내 식물체 분화)

  • Han, Young-Hee;Choi, Byoung-Ryourl;Soh, Ho-Seob;Lee, Seong-Jae;Choi, Young-Jin;Kim, Se-Young
    • Horticultural Science & Technology
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    • v.18 no.6
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    • pp.834-838
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    • 2000
  • As an endeavor to establish a micropropagation system for Epimedium koreanum Nakai., this study was carried out to define methods to disinfect its explants and media for callus induction, proliferation and plant regeneration. The lowest infection rates by fungi or bacteria on apical and axillary bud explants of rhizome were observed when they were immerged in 0.3% NaOCl solution for 20 min after soaked in 0.1% $AgNO_3$ solution for 30 min, but leaf explants were seldom infected with fungi or bacteria by this disinfectant method. The highest rate of plantlet formation was obtained from the explants disinfected in 0.3% NaOCl solution for 20 min after soaked in 0.1% $AgNO_3$ solution for 60 min for tip buds and in 0.1 % $AgNO_3$ solution for 30 min for axillary buds of rhizome. Induction rate of callus was the highest from the explants disinfectd in 0.3% NaOCl solution for 20 min after soaked in 0.2% $AgNO_3$ solution for 15 min. Callus growth was proper in a modified 1/2 MS medium including half strength of $NH_4NO_3$ with $0.02-0.2mg{\cdot}L^{-1}$ BA and $2.0mg{\cdot}L^{-1}$ NAA. Low rate of plantlet regeneration was obtained in 1/2 UM or 1/2 White medium with $2.0mg{\cdot}L^{-1}$ BA and $0.2mg{\cdot}L^{-1}$ AA.

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A Study on the Screening of the Novel Genes Associated with Lysosomal Trafficking and Mutation Detection in Fibroblasts of the Patients with Mucolipidosis type II and III (리소좀 교통 이상을 초래하는 뮤코지방증 2형과 3형 환자의 섬유아세포를 이용한 신규 유전자 탐색 및 돌연변이에 대한 연구)

  • Song, Seng Mi;Chang, Soo Hee;Paik, Kyung Hoon;Jin, Dong-Kyu
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.5 no.1
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    • pp.65-75
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    • 2005
  • Purpose: To understand genetic differences and similarities between mucolipidosis and control. Methods: Using the fibroblast of the mucolipidosis II and control, forward and reverse subtracted libraries were constructed. Among these clones, we investigated mutations in the GNPTA (MGC4170) gene, which codes for the ${\alpha}/{\beta}$ subunits of phosphotransferase, and in the GNPTAG gene, which codes for the ${\gamma}$ subunits in 5 Korean patients with mucolipidosis type II or IIIA. Result: Several differentially expressed cDNAs were cloned and their sequences were determined. Mutation analysis of the interested gene, GNPTA was performed and we identified 7 mutations in the GNPTA gene, but none in the GNPTAG gene. The mutations in type II patients included p.Q104X(c.310C>T), p.R1189X(c.3565C>T), p.S1058X(c.3173C>G), p.W894X(c.2681G>A) and p.H1158fsX15(c.3474_3475delTA), all of which are non-sense or frame shift mutations. However, a splicing site mutation, IVS13+1G>A (c.2715+1G>A) was detected along with a non-sense or a frame shift mutation (p.R1189X or p.E858fsX3(c.2574_2575delGA)) in two mucolipidosis type IIIA patients. Conclusion: This report shows that mutations in the GNPTA gene coding for the ${\alpha}{\beta}$subunits of phosphotransferase, and not mutations in the GNPTAG gene, account for most of mutations found in Korean patients with mucolipidosis type II or IIIA.

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