• Title/Summary/Keyword: agar-free

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Cloning of nif Gene Cluster from Klebsiella pneumoniae and Expression in Escherichia coli (Klebsiella pneumoniae의 nif Gene Cluster의 Cloning 및 Escherichia coli 에서의 발현)

  • 이재선;이성희;심웅섭
    • Korean Journal of Microbiology
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    • v.27 no.1
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    • pp.21-26
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    • 1989
  • The chromosomal DNA of Klebsiella penumoniae KNF1 was partially digested with HindIII. pBR322 was completely digested with same enzyme with additional alkaline phosphatase treatment. Both DNA samples were ligated and transformed into E. coli KO60. Single coloneis of the transformed cells were isolated on N0free agar media. These colonies were ampicillin-resistant and tetracycline-sensitive. When the plasmids in transformants were cured, nitrogen-fixing activities were lost. Therefore, these transformants harbored recombinant plasmid including nif genes of K. pneumoniae. Nitrogenase activity of transformant was higher than or the same as that of K. pneumoniae KNF1.

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Somatic Embryogenesis and Plant Regeneration from Poinsettia (Euphorbia pulcherrima L.) Stem Explants (포인세티아 줄기조직배양에 의한 재분화체 제조.)

  • Hee-Sung Park
    • Journal of Life Science
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    • v.8 no.6
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    • pp.623-626
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    • 1998
  • Conditions for somatic embryogenesis and plant regeneration from stem tissues of Euphorbia pulcherrima were esta-blished. Explants from leaf, petiole, stem were examined for their embryogenesis on MS solid medium supplemented with plant growth hormones in combination at various concentrations. From leaf or petiole explants, callus was indu-ced well but never proceeded to the embryonic stage in our expermental conditions. From stem explants, however, multiple shoots following callus induction emerged in about 6 to 8 weeks on MS agar medium supplemented with 1.5 mg/L of benzyladenine. Upon transfer, roots were developed on hormone-free MS solid medium.

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Antibacterial Activities of Lactobacillus crispatus ATCC 33820 and Lactobacillus gassed ATCC 33323

  • Kim, Jin-Woo;S.N. Rajagopal
    • Journal of Microbiology
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    • v.39 no.2
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    • pp.146-148
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    • 2001
  • Lactobacillus crispatus ATCC 33820 and L. gasseri ATCC 33323 were grown in MRS broth (pH 6.5) at 37$^{\circ}C$ for 24 h and the antibacterial activities of cell free culture supernatants were determined by the agar well diffusion method. The culture supernatants were inhibitory to Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Bacillus subtilis, Staphylococcus aureus, Enterococcus faecalis, Pediococcus acidilacticii, and Lactobacillus helveticus. The supernatants did not show any lysozyme activity. Addition of catalase did not affect the antibacterial activities of the supernatants. The antibacterial substances were heat stable (100$^{\circ}C$ for 60 min) and sensitive to proteases.

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Antibacterial Activity of Lactobacillus sakei BKl9 against Fish Pathogenic Bacteria (Lactobacillus sakei BK19의 어류 병원성 세균에 대한 항균활성)

  • 양병규;이제희;허문수
    • Korean Journal of Microbiology
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    • v.39 no.1
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    • pp.56-61
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    • 2003
  • The purpose of the present study was to screen the effective of lactic acid bacteria (LAB), as probiotics which are able to protect bacterial fish diseases and investigate their characteristics. Twenty strains of lactic acid bacteria were isolated from fish intestine. fermented fish foods and kimchis. These bacteria were screened for antagonistic activity against fish pathogenic bacteria. Seven tested LAB strains were able to inhibit the fish pathogenic bacteria, including Vibrio anguillarum, Edwardsiella tarda and Streptococcus sp. Of the probiotic candidates, BK19 strain which from fermented pollack viscera indicated the largest inhibition activity. This particular probiotic bacteria was identified and named as Lactobacillus sakei BK19. In the scanning electron microscope observation, L. sakei BK19 supernatant treated V.anguillarum cell wall had been destroyed incubate after 3 hr.

Effects of Storage Time on Bacteria Concentration and Sperm Parameters in Boar Semen (돼지 액상정액 보존 일수에 따른 정액내 세균과 정자 기능의 변화)

  • Chung, Ki-Hwa;Kim, In-Cheol
    • Reproductive and Developmental Biology
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    • v.36 no.3
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    • pp.163-166
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    • 2012
  • This study was conducted to determine the relationship between elapsed time after semen preservation on the changes of bacteria and semen quality. Semen was diluted with BTS(Beltsville Thawing Solution) extender without antibiotic for 7 days and sperm parameter and fertility were measured. Sperm motility was measured by CASA and total bacteria number was counted after 22~24 hr incubation from counting agar plate in which sperm dilute to $10{\sim}10^6$ in 0.9% saline solution and inoculate to agar. Acrosomal integrity was measured by Chlortetracycline (CTC) staining. CTC patterns were uniform fluorescence over the whole head (pattern F), characteristic of incapacitated acrosome-intact spermatozoa; fluorescence-free band in the post-acrosomal region (pattern B), characteristic of capacitated acrosome-intact spermatozoa; and almost no fluorescence over the whole head except for a thin band in the equatorial segment (pattern AR), characteristic of acrosome reacted spermatozoa. Total number of bacteria was significantly increased (p<0.0001) 3 days after preservation. Sperm motility, viability, and morphological abnormality on elapsed time after preservation were lower from 5 ($77.24{\pm}6.47$, p<0.001) and 7 days ($77.24{\pm}6.47$, p<0.001) after preservation compared to 1 ($15.71{\pm}7.18$) and 3 days($18.39{\pm}7.22$) after preservation, respectively. Sperm viability was significantly lower ($53.25{\pm}35.03$, p<0.0001) at 7 days after preservation. Morphological abnormality of sperm was lower (p<0.001) at 1 ($15.71{\pm}7.18$) and 3 ($18.39{\pm}7.22$) days compared to 5 ($21.84{\pm}7.91$) and 7 ($22.59{\pm}9.93$) days after preservation. Acrosomal integrity and capacitation rate (pattern F) were significantly lower (p<0.001) from 5 days after preservation. Based on the data we obtained from this study suggested that semen preserved more than 5 days without antibiotic would not recommend use for artificial insemination.

Estimation of Antibacterial Properties of Chlorophyta, Rhodophyta and Haptophyta Microalgae Species

  • Imran Bashir, Khawaja Muhammad;Lee, Jae-Hyeon;Petermann, Maike Julia;Shah, Abid Ali;Jeong, Su-Jin;Kim, Moo-Sang;Park, Nam-Gyu;Cho, Man-Gi
    • Microbiology and Biotechnology Letters
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    • v.46 no.3
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    • pp.225-233
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    • 2018
  • In this exploratory study, eight types of microalgae from different phyla (Chlamydomonas reinhardtii, Chlorella species, Haematococcus pluvialis, Porphyridium purpureum, Porphyridium cruentum, Isochrysis species, Isochrysis galbana, and Pavlova lutheri) were tested for their antibacterial activities against eight target pathogenic bacterial strains. The agar well diffusion method and broth micro dilution assay were conducted to estimate the antibacterial activity. Microalgae cell-free supernatants, exopolysaccharides (EPS), water, and organic solvent extracts were used for inhibition analysis. EPS extracted from P. lutheri showed activity against Bacillus subtilis and Pseudomonas aeruginosa. Inhibition zone diameters of 14-20 mm were recorded on agar plates, while the minimum inhibitory concentrations in the broth micro dilution assay were $0.39-25mg\;ml^{-1}$. During this study, haptophyte microalgae, Isochrysis species, and P. lutheri extracts showed the highest activity against most of the tested pathogenic bacterial strains, while most of the extracts were active against the important foodborne pathogen P. aeruginosa. This study showed promising results regarding important microalgae phyla, which will further aid research related to extracts and exploitation of bioactive metabolic compounds in the food and pharmaceutical industries.

Antifungal and Antioxidative Activities of Yucca smallina Fern

  • Jin, Yu-Lan;Jung, Woo-Jin;Kuk, Ju-Hee;Kim, Jung-Bong;Kim, Kil-Yong;Park, Ro-Dong
    • Journal of Applied Biological Chemistry
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    • v.49 no.4
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    • pp.165-170
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    • 2006
  • The antifungal activity of crude methanolic extract and fractions from Yucca smalliana Fern. leaves, roots and flowers were investigated in vitro against a panel of plant pathogenic fungi. The minimal inhibitory concentration(MIC) was determined by an agar dilution method. Preliminary liquid culture and agar plate assays showed that the growth of Fu sarium oxysporum, Phytophthora capsici, Rhizoctonia solani and Botrytis cinerea were inhibited by Y. smalliana extracts. The extracts from flowers and leaves showed antifungal activity of 64.0% and 34.0% against F. oxysporum, 66.0% and 62.0% against P. capsici, and 27.0% and 41.0% against B. cinerea, respectively. The methanolic extract from Y. smallina leaves in distilled water was fractionated using solvents of increasing polarity: hexane, ethyl acetate and butanol. These fractions had a broad spectrum of antifungal activity, found to reside entirely in the butanol and aqueous fraction. The aqueous fraction showed inhibition rate of 60.0, 67.8, 84.6 and 58.3% against F. oxysporum, R. solani, C. gloeosporioides, and B. cinerea, respectively, and the butganol fracgtion showed 36.0, 46.0, 66.1 and 58.3%, respectively. Phenolics(e.g. flavonoids, steroids and terpenoids) were observed in the thin layer profile of the different fractions. Leave extract showed a prominent antioxidant activity totally scavenging the free radical of DPPH at a concentration of 1 mg/ml.

Efficient Liquid Media for Encystation of Pathogenic Free-Living Amoebae

  • Sohn, Hae-Jin;Kang, Heekyoung;Seo, Ga-Eun;Kim, Jong-Hyun;Jung, Suk-Yul;Shin, Ho-Joon
    • Parasites, Hosts and Diseases
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    • v.55 no.3
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    • pp.233-238
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    • 2017
  • Pathogenic Naegleria fowleri, Acanthamoeba castellanii, and Acanthamoeba polyphaga, are distributed worldwide. They are causative agents of primary amoebic meningoencephalitis or acanthamoebic keratitis in humans, respectively. Trophozoites encyst in unfavorable environments, such as exhausted food supply and desiccation. Until recently, the method of N. fowleri encystation used solid non-nutrient agar medium supplemented with heat-inactivated Escherichia coli; however, for the amoebic encystment of Acanthamoeba spp., a defined, slightly modified liquid media is used. In this study, in order to generate pure N. fowleri cysts, a liquid encystment medium (buffer 1) modified from Page's amoeba saline was applied for encystation of N. fowleri. N. fowleri cysts were well induced after 24 hr with the above defined liquid encystment medium (buffer 1). This was confirmed by observation of a high expression of differential mRNA of nfa1 and actin genes in trophozoites. Thus, this liquid medium can replace the earlier non-nutrient agar medium for obtaining pure N. fowleri cysts. In addition, for cyst formation of Acanthamoeba spp., buffer 2 (adjusted to pH 9.0) was the more efficient medium. To summarize, these liquid encystment media may be useful for further studies which require axenic and pure amoebic cysts.

Somatic Embryogenesis and Plant Regeneration in Immature Flower Bud Cultures of Carnation (카네이션의 미숙화뢰 배양을 통한 체세포배 발생 및 식물체 재분화)

  • 안병준
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.6
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    • pp.369-374
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    • 1997
  • Immature flower buds of 'Desio' carnation were cultured on MS agar medium supplemented with 1 ㎎/L 2,L-D. Embryogenic calli were formed from 5-10% of the buds less than 20 ㎜ in length, but only non-embryogenic calli were produced from explants of shoot apex leaf, internode, and flowere buds larger than 20 ㎜. The same method was applied to 16 cultivars of cut Sower carnation and embryogenic calli were obtained in 7 cultivars. Several embryogenic callus lines were selected and maintained through subcultures over 120 weeks without loss of embryogenic competence. The embryogenic cultures were also proliferated rapidly in liquid agitation cultures using MS medium supplemented with 1mg/L 2,4-D. Numerous embryos were formed on the periphery of the cell aggregates upon transfer to auxin-free MS agar medium. Plantlets were transplanted in potting soil and grown to bloom in six months.

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Quality Characteristics of Yanggeng added with Pesticide Free Wild Grape (Vitis amurensis) Juice (무농약 산머루 즙을 첨가한 양갱의 품질특성)

  • Kang, Chang-Soo
    • Korean Journal of Organic Agriculture
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    • v.23 no.2
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    • pp.301-309
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    • 2015
  • This study was performed to investigate the quality characteristics of Yanggeng added wild grape (Vitis amurensis) juice. The yanggeng was prepared with pesticide-free wild grape juice, agar, sugar and salt. The yanggeng was made with various levels (0, 50, 100, 150, 200 g of wild grape juice in yanggeng, respectively) based on the total weight of water. It was estimated on Hunter's color value, texture profile analysis, and sensory characteristics of the Yanggeng. As the content of wild grape juice increased, The lightness (L) and yellowness (b) decreased and redness (a) increased. In texture profile analysis, hardness was increased; however, springiness, cohesiveness, gumminess and chewiness decreased with increasing levels of wild grape juice. The results of the sensory evaluation indicated that the Yanggeng containing the 150 g level of wild grape juice showed the highest preference scores in terms of color, taste, texture, flavor and overall acceptance. These results show that the yanggeng containing 150 g of wild grape juice is the better.