• 제목/요약/키워드: in vitro cultivation

검색결과 189건 처리시간 0.028초

신경괴사증바이러스(Nervous Necrosis Virus, NNV) 모니터링을 통한 무감염 능성어(Hyporthodus septemfasciatus)친어의 선발 (Monitoring of Nervous Necrosis Virus (NNV) in the Broodstock of Seven Band Grouper Hyporthodus septemfasciatus)

  • 김시우;김위식;서한길;김경민;오명주
    • 한국수산과학회지
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    • 제50권5호
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    • pp.527-533
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    • 2017
  • We investigated the infection of nervous necrosis virus (NNV) in seven band grouper Hyporthodus septemfasciatus broodstocks, which have been reared in aquaculture farms in South Korea during 2012-2014. To investigate the prevalence of NNV within the broodstock, egg, sperm, and blood were sampled in the spawning season. The egg and sperm samples were subjected to a nested reverse transcription (RT) polymerase chain reaction (PCR) assay to detect NNV and were inoculated on SSN-1 cells to culture the virus. Blood samples were used to detect antibodies against NNV using enzyme linked immunosorbent assay (ELSIA). Positive values from ELISA were found in 39 of 162 samples (24%) in 2012, and 13 of 28 samples (46%) in 2014. Additionally, 4 of 34 broodstocks (11%) investigated in 2013-2014 were determined to be carriers from the nested RT-PCR and in vitro cultivation. The broodstocks in which antibodies against NNV were detected by ELISA, or in which NNV was detected by the nested RT-PCR assay, posed a risk of vertical transmission of NNV. Therefore, it is necessary to select virus-free broodstocks in seed production to reduce the possibility of the vertical transmission of NNV.

Viability and Functions of Alginate-microencapsulated Islets Isolated from Neonatal Pigs

  • Lin, Yi-Juain;Wang, Jui-Ping;Chung, Yu-Tung;Sun, Yu-Ling;Chou, Yu-Chi
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권5호
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    • pp.795-801
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    • 2007
  • Patients with Type I diabetes mellitus have been treated with porcine insulin for several decades and pigs have recently been deemed an ideal source of microencapsulated islet cells for clinical xenotransplantation. In this study, neonatal pigs were anesthetized and sacrificed prior to a pancreatectomy. Islet cells were isolated from pancreas via collagenase digestion. Islet cells were separated and collected by hand under microscopic guidance. These cells were suspended in 1.4% sodium alginate solution and encapsulated by dropping them into 1.1% calcium chloride solution and in which the round gel in size was 250-400 ${\mu}m$ in diameter. Viability of the microencapsulated islet cells cultured in medium at $37^{\circ}C$ was assessed by MTT assay. Furthermore, insulin released in response to glucose challenge was investigated using an enzyme-linked immunosorbent assay. Secretion of insulin was low in response to the basal glucose solution (4.4 mM) in medium and was significantly higher in response to the high glucose solution (16.7 mM). The viability of microencapsulated islet cells did not differ significantly over a period of 7 days; that is, the increasing pattern of insulin concentration in the culture medium after glucose stimulation interval day was similar throughout the 7 days cultivation. In summary, experimental evidences indicated that the effects of alginate-microencapsulation prolonged survival of the neonatal porcine islets in vitro cultures and the insulin response to glucose of the islets was maintained.

Changes in Gut Microbial Community of Pig Feces in Response to Different Dietary Animal Protein Media

  • Jeong, Yujeong;Park, Jongbin;Kim, Eun Bae
    • Journal of Microbiology and Biotechnology
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    • 제30권9호
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    • pp.1321-1334
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    • 2020
  • Beef, pork, chicken and milk are considered representative protein sources in the human diet. Since the digestion of protein is important, the role of intestinal microflora is also important. Despite this, the pure effects of meat and milk intake on the microbiome are yet to be fully elucidated. To evaluate the effect of beef, pork, chicken and milk on intestinal microflora, we observed changes in the microbiome in response to different types of dietary animal proteins in vitro. Feces were collected from five 6-week-old pigs. The suspensions were pooled and inoculated into four different media containing beef, pork, chicken, or skim milk powder in distilled water. Changes in microbial communities were analyzed using 16S rRNA sequencing. The feces alone had the highest microbial alpha diversity. Among the treatment groups, beef showed the highest microbial diversity, followed by pork, chicken, and milk. The three dominant phyla were Proteobacteria, Firmicutes, and Bacteroidetes in all the groups. The most abundant genera in beef, pork, and chicken were Rummeliibacillus, Clostridium, and Phascolarctobacterium, whereas milk was enriched with Streptococcus, Lactobacillus, and Enterococcus. Aerobic bacteria decreased while anaerobic and facultative anaerobic bacteria increased in protein-rich nutrients. Functional gene groups were found to be over-represented in protein-rich nutrients. Our results provide baseline information for understanding the roles of dietary animal proteins in reshaping the gut microbiome. Furthermore, growth-promotion by specific species/genus may be used as a cultivation tool for uncultured gut microorganisms.

마렉병 바이러스 감염과 병원성 발현 기전 (Infection and Pathogenesis Mechanisms of Marek's Disease Virus)

  • 장형관;박영명;차세연;박종범
    • 한국가금학회지
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    • 제35권1호
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    • pp.39-55
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    • 2008
  • Like the other herpesviruses, the virion of MDV consists of an envelope, which surrounds an amorphous tegument. Within the tegument, and icosahedral capsid encloses a linear double-stranded DNA core. Although the genome structure of MDV indicates that it is an ${\alpha}-herpesvirus$ like herpes simplex and varicella-zoster viruses, biological properties indicate MDV is more akin to the ${\gamma}-herpesvirus$ group, which includes Epstein-Barr and Kaposi's sarcoma herpesviruses. These herpesviruses replicate lytically in lymphocytes, epithelial and fibroblastic cells, and persist in lymphoblastoid cells. MDV has a complex life cycle and uses two means of replication, productive and non-productive, to exist and propagate. The method of reproduction changes according to a defined pattern depending on changes in virus-cell interactions at different stages of the disease, and in different tissues. Productive (lytic) interactions involve active invasion and take-over of the host cell, resulting in the production of infectious progeny virions. However, some herpesviruses, including MDV, can also establish a non-productive (abortive) infection in certain cell types, resulting in production of cell-associated progeny virus. Non-productive interactions represent persistent infection, in which the viral genome is present but gene expression is limited, there is no structural or regulatory gene translation, no replication, no release of progeny virions and no cell death. Reactivation of the virus is rare, and usually the infectious virus can be re-isolated only after cultivation in vitro. MDV establishes latency in lymphoid cells, some of which are subsequently transformed. In this review article, recent knowledges of the pathogenesis mechanisms followed by MDV infection to sensitive cells and chickens are discussed precisely.

Alteration of cellular events in tooth development by chemical chaperon, Tauroursodeoxycholic acid treatment

  • Lee, Eui-Seon;Aryal, Yam Prasad;Kim, Tae-Young;Pokharel, Elina;Kim, Harim;Sung, Shijin;Sohn, Wern-Joo;Lee, Youngkyun;An, Chang-Hyeon;Kim, Jae-Young
    • International Journal of Oral Biology
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    • 제45권4호
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    • pp.190-196
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    • 2020
  • Several factors, including genetic and environmental insults, impede protein folding and secretion in the endoplasmic reticulum (ER). Accumulation of unfolded or mis-folded protein in the ER manifests as ER stress. To cope with this morbid condition of the ER, recent data has suggested that the intracellular event of an unfolded protein response plays a critical role in managing the secretory load and maintaining proteostasis in the ER. Tauroursodeoxycholic acid (TUDCA) is a chemical chaperone and hydrophilic bile acid that is known to inhibit apoptosis by attenuating ER stress. Numerous studies have revealed that TUDCA affects hepatic diseases, obesity, and inflammatory illnesses. Recently, molecular regulation of ER stress in tooth development, especially during the secretory stage, has been studied. Therefore, in this study, we examined the developmental role of ER stress regulation in tooth morphogenesis using in vitro organ cultivation methods with a chemical chaperone treatment, TUDCA. Altered cellular events including proliferation, apoptosis, and dentinogenesis were examined using immunostaining and terminal deoxynucleotidyl transferase dUTP nick end labeling assay. In addition, altered localization patterns of the formation of hard tissue matrices related to molecules, including amelogenin and nestin, were examined to assess their morphological changes. Based on our findings, modulating the role of the chemical chaperone TUDCA in tooth morphogenesis, especially through the modulation of cellular proliferation and apoptosis, could be applied as a supporting data for tooth regeneration for future studies.

에틸렌 저해제에 의한 배추 자엽조직의 기내 재분화율 향상 (Enhancement of Shoot Regeneration by Ethylene Inhibitors from Cotyledon Explant of Brassica campestris L. ssp. pekinensis)

  • 이혜승;조화진;김병동
    • 식물조직배양학회지
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    • 제22권5호
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    • pp.267-271
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    • 1995
  • 배추의 기내재분화율을 향상시키기 위하여 자엽을 치상재료로 하여 에틸렌 억제제인 AgNO$_3$와 STS를 첨가하여 그 영향을 조사하였다. AgNO$_3$의 경우에는 100 $\mu$M이 첨가되었을 때 STS에서는 5 $\mu$M이 첨가되었을 때, 무처리구의 4%, 2%에 비해 각각 가장 높은 22%와 15.3%의 재분화율을 보였다. 자엽연령에 대해서는 파종 후 3일 지난 자엽을 사용하였을 때 58%의 재분화율을 나타내었다. 배양용기에 따른 효과는 1회용 페트리디쉬에서 가장 높았으나 처리간에 뚜렷한 차이는 없었다. 뿌리유기 조건은 생장조절제가 포함되지 않은 처리와 NAA가 0.1 mg/L 농도로 첨가된 배지에서 가장 좋은 결과를 얻었으며 그 형태 및 밀도, 활력에서 가장 우수하였다. 기내에서 뿌리가 형성된 식물체를 4$^{\circ}C$에서 4주간 처리하여 추대를 유기하였고 형태적으로 정상적인 화기를 볼 수 있었으며 이들로부터 종자를 얻을 수 있었다.

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Evaluation of Nematicidal Activity of Streptomyces yatensis KRA-28 against Meloidogyne incognita

  • Park, Eun-Jae;Jang, Hyun-Jae;Park, Chan Sun;Lee, Seung-Jae;Lee, Soyoung;Kim, Kang-Hoon;Yun, Bong-Sik;Lee, Seung Woong;Rho, Mun-Chual
    • Journal of Microbiology and Biotechnology
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    • 제30권5호
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    • pp.700-707
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    • 2020
  • The root-knot nematode (Meloidogyne incognita) is an important pathogen in crop cultivation, however, few methods are available to control this parasitic roundworm. In this study, the nematicidal effects of approximately 30 Streptomyces strains isolated from soil samples of Mt. Naejang (Korea) were tested against Meloidogyne incognita, and the culture broth of the strains KRA-24 and KRA-28 exhibited approximately 75% and 85% insecticidal activity, respectively, in in vitro assays. In in vivo pot experiments, these strains reduced the number of nematodes in the soil and the number of egg masses in the roots of red peppers. The two strains also survived in the presence of insecticidal agents (0.1 to 3.0%) such as fosthiazate, ethoprophos and terbufos when they were used in parallel. The mixture of KRA-24 or KRA-28 culture broth and fosthiazate exhibited nematicidal effects that were similar to those observed when KRA-24 or KRA-28 were used alone. Our results clearly suggest that the Streptomyces strains KRA-24 and KRA-28 should be promoted as a biocontrol agent against Meloidogyne incognita.

Integration and Expression of Goat ${\beta}-Casein/hGH$ Hybrid Gene in a Transgenic Goat

  • Lee, Chul-Sang;Lee, Doo-Soo;Fang, Nan-Zhu;Oh, Keon-Bong;Shin, Sang-Tae;Lee, Kyung-Kwang
    • Reproductive and Developmental Biology
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    • 제30권4호
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    • pp.293-299
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    • 2006
  • In order to generate transgenic goats expressing human growth hormone (hGH) in their mammary glands, goat ${\beta}-Casein/hGH$ hybrid gene was introduced into goat zygotes by pronuclear microinjection. DNA-injected embryos were transferred to the oviduct of recipients at 2-cell stage or to the uterus at morula/blastocyst stage after cultivation in glutathione-supplemented mSOF medium in vitro. Pregnancy and survival rate were not significantly different between 2-cell embryos and morula/blastocysts transferred to oviduct and uterus, respectively. One transgenic female goat was generated from 153 embryos survived from DNA injection. Southern blot analysis revealed that the transgenic goat harbored single-copy transgene with a partial deletion in its sequences. Despite of the partial sequence deletion, the transgene was successfully expressed hGH at the level of $72.1{\pm}15.1{\mu}g/ml$ in milk throughout lactation period, suggesting that the sequence deletion had occurred in non-essential part of the transgene for the transgene expression. Unfortunately, however, the transgene was not transmitted to her offspring during three successive breeding seasons. These results demonstrated that goat ${\beta}-casein/hGH$ gene was integrated into the transgenic goat genome in a mosaic fashion with a partial sequence deletion, which could result in a low level expression of hGH and a failure of transgene transmission.

주걱송편버섯(Pycnoporus cinnabarinus SCH-3)의 Laccase에 의한 Acridine 산화 (Oxidation of Acridine by Laccase of Pycnoporus cinnabarinus SCH-3)

  • 이현수;한만덕;윤경하
    • 미생물학회지
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    • 제44권2호
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    • pp.110-115
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    • 2008
  • Acridine은 fungal laccase의 기질이 아님에도 불구하고 acridine을 Pycnoporus cinnabarinus SCH-3 배양액에 첨가했을 때 acridone으로 산화되었다. P. cinnabarinus SCH-3균주는 배양 중에 다량의 laccase와 3-hydroxyanthranilic acid (3-HAA)와 cinnabarinic acid (CA)를 생성했다. 정제된 laccase와 acridine을 완충용액에서 직접 반응시켰을 때 acridine은 변화되지 않았다. 그러나 laccase의 기질인 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS)나 3-HAA를 laccase와 acridine 혼합액에 첨가했을 경우에는 acridine이 acridone으로 산화되었다. 특히 ABTS 첨가구는 3-HAA 첨가구보다 acridine 산화율이 2배 이상 높았다. 한편 정제된 laccase와 3-HAA를 완충액에서 반응시켰을 때 3-HAA는 CA로 전환되었다. 이와 같은 실험결과들은 P. cinnabarinus SCH-3의 laccase가 배양중에 생산된 3-HAA를 매개체로 사용하여 acridine을 acridone으로 산화하고 CA는 laccase에 의하여 3-HAA로부터 합성됨을 나타낸다.

농업미생물은행(KACC)의 곰팡이 보존 (Maintenance of Filamentous Fungi in Korean Agricultural Culture Collection (KACC))

  • 신명숙;홍승범
    • 한국균학회지
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    • 제42권2호
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    • pp.97-103
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    • 2014
  • 농업미생물은행(KACC)에는 7,039균주의 곰팡이가 장기 보존되어 있다. 이 중에서 다수의 포자를 형성하는 4,065균주는 동결건조법으로 보존되어 있는데 사멸로 인한 균주의 소실을 막기 위하여 대부분은 액체질소보존법 그리고 모든 균주가 저온냉동고보존법으로 함께 보존되어 있다. 4,065균주의 곰팡이에는 주로 공기 중에 흔하며 산업적으로 이용성이 많은 Aspergillus, Penicillium, Lichtheimia, Mucor, Rhizopus 속 등이 포함된다. 포자를 형성하지 않거나 소수형성하거나 또한 보존이 어려운 너무 큰 포자를 형성하는 나머지 곰팡이는 동결건조보존법으로 보존이 불가하기 때문에 액체질소보존법, 저온냉동고보존법 그리고 광유보존법으로 균주를 보존한다. 여기에는 Phytophthora, Pythium, Cercospora, Septoria, Rhizoctonia 속 등의 식물병원균이 포함된다. 이 외에도 KACC에서 이용하는 다양한 곰팡이 보존법을 소개하고 이들의 상세보존 과정을 기술한다.