• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.72-72
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• 2002

조류는 발생학적 특성상 수정과 초기 배발생을 제외한 거의 대부분의 개체발생 과정이 난각 속에서 진행된다. 그러므로 수정란에 생명 공학적 기법을 적용하는데 있어서, 포유류의 경우 여러 생명공학 기법이 적용된 수정란은 초기발생을 위한 체외배양 이후 반드시 모체에 이식되어야 하지만, 조류의 경우 인공적인 체외배양 체계가 확립되어 있어야 생명공학 기법이 적용된 수정란을 개체까지 발생시킬 수 있게 된다. 닭의 난자는 난관 누두부에 배란 후 약 15분내에 정자의 침입을 받아 수정되어 난관 팽대부에 도달하면 1세포기 수정란이 된다. 그 후 수정란이 협부에 도달하면 최초로 분할이 일어나기 시작하여, 방란시에는 그 세포수가 60,000개에 이르게 된다. 한편, 계란의 형성 과정에서는 다량의 난황을 포함한 난자가 난관 누두부로 배란되어 정자와 만나게 되면 수정란으로서 계란 형성이 계속되고 정자와 만나지 못하게 되면 무정란으로서 계란 형성을 계속하게된다. 배란된 난자가 난관누두부를 거쳐 난관팽대부에 도달되면 난자는 농후난백에 의해 둘러싸이게 되고 난관혈부에 도달되면 난각막이 형성되고 수양성 난백이 침적하게 된다. 그 후, 난관협부를 지난 난자는 난관자궁부에 도달되면 20시간이상 그곳에 머물면서 난각형성이 진행된 다음 방란된다. 따라서 수정란에 외래유전자를 미세주입하여 형질전환 닭을 생산하기 위해서는 수정란을 암탉의 난관 내에서 최초 분할되기 전에 외부로 끄집어내어야 하며, 수정란에 외래 유전자를 미세주입한 다음에는 다시 암탉의 난관내로 이식해야 하지만 현재까지 그러한 기술은 확립되어 있지 못하다. 그렇기 때문에 모체의 난관 속에서 일어나는 배 발생과 그 이후 개체까지의 발생을 위하여 인공적인 체외배양 체계가 확립되어 있어야 한다. 따라서 본 발표에서는 형질전환 닭을 생산하기 위한 양질의 1세포기 수정란 획득 방법과 획득된 수정란의 체외 배양방법에 관하여 기술적인 측면에서 고찰 해보고자 하며, 그와 같은 배양 기술을 이용하여 외래유전자를 도입한 일련의 결과에 관하여 보고 하고자한다.

• FLOWER RESEARCH JOURNAL
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• v.17 no.2
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• pp.114-120
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• 2009

The most effective conditions of in vitro culture were studied for mass propagation of Pteris cretica 'Wilsonii'. Spores of the species germinated within 7 weeks. The greatest proliferation was obtained with Knop and Hyponex media, but growth was more effective in Hyponex medium. MS medium induced necrosis of prothalli in all strength of nitrogen and sucrose except in case of 0% sucrose. Hyponex medium supplemented with 1% sucrose and 0.6% agar promoted propagation and growth of prothalli. In Hyponex medium, optimal inoculation method was homogenization, but in MS medium dividing colonies of prothalli was more effective. Culturing on solid medium was more effective than liquid culture method. Liquid culture induced necrosis of prothalli. Shaking cultured prothalli showed good growth, but propagation was inhibited compared to those cultured on solid medium.

• The Korean Journal of Mycology
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• v.47 no.2
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• pp.153-163
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• 2019

This study was carried out to investigate the feasibility of Salix gracilistyla production for cosmetic use through mycelial fermentation. The efficacy of this method was confirmed by fermentation using the mycelia of Ganoderma lucidum (a representative medicinal mushroom). Total polyphenol and flavonoid content and DPPH radical scavenging activity of S. gracilistyla extract (SGE) were found to be higher than those of G. lucidum fermented S. gracilistyla extract (GLSGE). GLSGE had relatively lower collagenase activity than SGE. However, GLSGE increased HDFn cell viability more potently than SGE, and increased the biosynthesis of type I procollagen. Thus, GLSGE could be used as an anti-aging cosmetic active ingredient. These results indicate that extract fermentation using G. lucidum mycelia can effectively enhance some beneficial effects of functional materials.

• Korean Journal of Clinical Laboratory Science
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• v.47 no.4
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• pp.313-317
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• 2015

Due to the increase in incidence of infection of Mycobacterium tuberculosis complex (MTC), it is imperative that a rapid diagnosis accompanies the handling of MTC. This is due to the three to eight weeks it takes to culture Mycobacteria, and the lack of sensitivity of microscopic examination of AFB. Recently, nested PCR has been used to detect and diagnose mycobacteria. It is especially useful in complementing diagnosis by histological extra pulmonary. After culturing all the specimens and practicing the nested PCR, we did comparison analysis between nested PCR and culture. There were 76 specimens, 31 of which were positive. Of the 31 positive specimens in culturing, only 22 were positive in nested PCR. Of the 45 negative specimens, 36 were negative in nested PCR. As a result, Sensitivity was 71% and specificity was 80%. Furthermore, the positive predictive value was 71% and negative predictive value was 80%. These results indicate that nested PCR based techniques are sensitive, specific, and rapid methods for the detection of MTC.

• KSBB Journal
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• v.18 no.3
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• pp.197-201
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• 2003

Automatic addition of glucose, ammonium and phosphate to alcohol distillery wastewater and their control at low concentrations have been carried increase the cell concentration of a baker's yeast cultivated in the wastewater. Glucose was automatically added using dissolved oxygen as the control parameter, and maintained below 300 mg/L. Ammonium was automatically added by a pH-stat method and maintained in the low range of 12.6~17.4 mM. An automated FIA system, which used an ascorbic acid-based method was developed for the automatic analysis nad addition of phosphate. With this system, the phosphate concentration was succesfully analysed and controlled afrer 19.4 hr in the range 23.3~43.4 mg/L. The cell concentration was increased by 33.0-fold by the addition of these three nutrients. The overall specific growth rate of the yeast was 0.19 $hr^{-1}$.

• Journal of Mushroom
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• v.19 no.2
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• pp.109-113
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• 2021

Currently, the spawn of the mushroom Agarcus bisporus is produced by a method developed in the 1980s, and anew spawning method needs to be developed to improve the quality of the spawn. In this study, the condition for a maximum mycelium weight(5.92±0.52 g/L) was shaking culture (24 hours/day) at 24℃ and 120 rpm in CDB (compost dextrose broth). Based on this, the ventilated liquid culture method (2.5 L/min) was cultured for 10 days. This method was appropriate, andwhen the inoculum was cultured at 50 g/mL for about 10 days, it was cultured well without agglomeration and shaking of seed.

• FLOWER RESEARCH JOURNAL
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• v.17 no.3
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• pp.172-178
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• 2009

This experiment was conducted to elucidate the proper suppling frequency of nutrient solution for grafted cactus Gymnocalycium mihanovichii var. friedrichii grown hydroponically without medium. Grafted cactus seedlings were planted onto the cultivation bed without medium, using labor-saving tray. The treatments like 1, 3, 5 and 7 times of nutrient solution supply per day and continuous soaking of plant root in the nutrient solution during the daytime were tested in summer and winter season. The growth of grafted cactus was worst in the treatment of one time supply of nutrient solution per day, and there were not significant difference in growth of grafted cactus among other treatments both in summer and winter season. 17.6% of grafted cactus seedlings failed to rooting in the treatment of one time supply of nutrient solution per day in winter season. The proper suppling frequency of nutrient solution, for the grafted cactus Gymnocalycium mihanovichii var. friedrichii grown hydroponically without medium, was three times supply of nutrient solution per day both in summer and winter seasons considering growth and rooting of plants.

• Tuberculosis and Respiratory Diseases
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• v.47 no.6
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• pp.747-756
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• 1999

Background: Acid-fast stain and cultures for diagnosis of pulmonary tuberculosis are primary and essential method, but have their limitation : low sensitivity and time consuming. The objective of this study is comparison of amplified Mycobacterium tuberculosis direct test(MTD) by the conventional AFB smears and cultures in the detection of Mycobacterium tuberculosis in respiratory specimens. Methods: During the period between November, 1997 and May, 1998 a total of 267 respiratory specimens (sputum 173, bronchial washing 94) from 187 patients suspected pulmonary tuberculosis were subjected to AFB smears, cultures and MID test. MID is based on nucleic acid amplification. We compared the MID with 3% Ogawa culture method. In positive AFB smear and negative MID specimen, positive culture identification between nontuberculous mycobacterium and M.tuberculosis was assesed by using Accuprobe M.tuberculosis complex probe. In negative AFB smear and negative AFB culture, MTD results are assessed by clinical follow-up. Results : 1) Compared with culture in sputum and bronchial fluid specimens, sensitivity and specificity of MTD in positive AFB smear is 79.7% and 20.0%, sensitivity and specificity of MTD in negative AFB smear specimens is 75.0% and 79.7%. 2) Discrepant analysis is assessed by clinical follow-up and other specimen results beyond study. Culture negative but MTD positive specimens were proved to be true positive and gave MTD sensitivity 79.2%, specificity of 84.4%, positive predictive value 80.5% and negative predictive value 83.2%. 3) 14 out of 31 specimens in negative AFB smear, negative AFB culture and positive MTD showed pulmonary tuberculosis diagnosed on clinical follow-up and sensitivity is 45.2%. 4) 2 out of 13 specimens in positive AFB smear, positive AFB culture and negative MID diagnosed as non tuberculous mycobacterium by Accuprobe culture. Conclusion: This study suggested that MID in respiratory specimens is simple and rapid diagnostic method, but considered adjuvant method rather than replace the conventional AFB smear and culture.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.347-353
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• 1997

This study was to test whether in vitro/in vivo survival of vitrified mouse blastocysts was influenced by culture conditions and ET method. Mouse blastocysts were obtained from in vitro fertilization and cultured for 4 days in M16 medium, and they were vitrified in EFS40 which contained 40% ethlyene glycol, 18% Ficoll and 0.5 mol sucrose in PBS. In experiment I, in vitro and in vivo survival rate of these embryos were evaluated in different culture condition after thawing. When thawed embryos were cultured in M16 medium as a control, m-CR1 medium contained 20 amino acids (2% BME amino acis and 1% MEM non-essential amino acids solution) and 4 mg/ml BSA and cumulus monolayer cell co-cultured condition in mCR1 medium (10% FBS), their in vitro survival at 24 hr after thawing was not affected by culture condition (75.6, 83.1, 82.4%). However, in vivo survival rates of implantation in m-CR1 medium (80.4%) were significantly higher than those of M16 medium (51.2%), co-culture (57.1%) condition, although there was no difference in live fetuses rates on day 15 gestation (39.0, 49.0, 38.1%). In experiment II, the in vivo development potential of embryos by ET methods was examined. When blastocysts were transferred to the day 2, 3 pseudopregnant recipient without culture soon after thawing, no pregnant recipient was obtained on the day 2 pseudopregnancy, and 50% of pregnancy rates and 15.4% of live fetus rates were obtained on the day 3 pseudopregnant recipients. These results were significantly lower than those of transferred group (day 3 pseudopregnant recipients) after culture for 16 hr post thawing (73.5, 57.1%) (p<0.05). In experiment III, to elevate usability of delayed embryos in vitro/in vivo survival of vitrified embryos (day 4 early, day 5 early and expanding blastocyst) were examined. in vivo survival rates (live fetus, total implantation) were higher in day 4 early blastocysts (33.3, 66.7%) than in day 5 expanding blastocysts (29.0, 38.7%), although the highest in vitro survival rates were obtained in the day 5 expanding brastocysts (78.3%). Therefore, these results suggest that the in vitro/in vivo survival rates of vitrified embryos could be improve by the culture condition and ET method and that the in vivo development rates of delayed embryos were decreased with longer culture duration in vitro. It means that more effective cryopreservation was obtained in day 4 early blastocysts than in day 5 expanding blastocysts.

• Microbiology and Biotechnology Letters
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• v.39 no.4
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• pp.374-381
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• 2011

The beneficial effects of Eisenia fetida on soil properties have been attributed to their interaction with soil microorganisms. The bacterial diversity of the intestinal tract of E. fetida was investigated by culture-dependent and culture-independent methods including denaturing gradient gel electrophoresis (DGGE) and pyrosequencing analyses. In a pure culture, Lysinibacillus fusiformis (51%), Bacillus cereus (30%), Enterobacter aerogenes (21%), and L. sphaericus (15%) were identified as the dominant microorganisms. In the DGGE analyses, B. cereus (15.1%), Enterobacter sp. (13.6%), an uncultured bacterium (13.1%), and B. stearothermophilus (7.8%) were identified as the dominant microorganisms. In the pyrosequencing analyses, Microbacterium soli (26%), B. cereus (10%), M. esteraromaticum (6%), and Frigoribacterium sp. (6%) were identified as the dominant microorganisms. The other strains identified were Aeromonas sp., Pseudomonas sp., Borrelia sp., Cellulosimicrobium sp., Klebsiella sp., and Leifsonia sp. The results illustrate that culture independent methods are better able to detect unculturable microorganisms and a wider range of species, as opposed to isolation by culture dependent methods.