• 제목/요약/키워드: Insertion strain

검색결과 88건 처리시간 0.025초

Prostaglandin $E_2$가 백서의 치아이동시 치조골 흡수에 미치는 영향에 관한 실험적 연구 (AN EXPERIMENTAL STUDY ON THE EFFECT OF PROSTAGLANDIN $E_2$ ON ALVEOLAR BONE RESORPTION INDUCED BY TOOTH MOVEMENT IN RATS)

  • 강봉기;서정훈
    • 대한치과교정학회지
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    • 제13권2호
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    • pp.147-154
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    • 1983
  • This experiment was performed to study the effect of $PGE_2$ on the bone resorption at the tooth movement by orthodontic force. The experimental animals were the Sprague-Dawley strain rats. The orthodontic force was applied by the insertion of separating clamp made of 0.014' (0.356mm) wire to the interproximal site between the 2nd and the 3rd upper right molars. In experiment I, $0.2{\mu}g,\;0.4{\mu}g,\;0.8{\mu}g,\;and\;1.0{\mu}g\;PGE_2$ were locally injected at the submucosa near the 2nd molar of the maxilla each. The effect was detected by the count of the number of osteoclasts appeared at the compressed surface of interradicular bone. In experiment II, 1.0 mg/kg indomethacin (a specificc inhibitor of prostaglandin synthetas.) was subcutaneously injected. The effect was examined by the count of the number of cateoclasts appeared at the compressed surface of interradicular bone. The obtained results were follows; 1. The number of osteoclasts on the compressed surface of the interradicular bone increased in proportion to the increased dosage of $PGE_2$ administered. The number of osteoclasts increased significantly at the administration of $0.8{\mu}g\;and\;1.0{\mu}g\;PGE_2$ in contrast to the control (P<0.05). 2. The administration of 1.0 mg/kg indomethacin decreased the number of osteoclasts at the compressed bony surface significantly (P<0.01).

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Bacillus sp. SSA3 균주의 Expression Vector 개발 (Construction of Expression Vector of Bacillus sp. SSA3 Strain)

  • 조윤래;김종규;권대준
    • 한국미생물·생명공학회지
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    • 제20권6호
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    • pp.637-641
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    • 1992
  • 한국 재래식 된장.간장 발효균 Bacillus sp. SSA3 균주의 expression vector를 개발하기 위해 Bacillus sp. SSA3의 chromosomal DNA로부터 유전자의 promoter 부위를 cloning 하였다. Recombinant plasmid를 제작하기 위하여 Bacillus sp. SSA3의 chromosomal DNA을 HindIII로 절단한 단편을 pGR71 plasmid의 CAT gene과 pUC18 plasmid의 $\beta$-galactosidase gene의 전방에 삽입시킨 후, E.coli JM109에 형질전환하였다.E. coli JM109의 chloramphenicol 내성 clones으로부터 6 recombinant plasmid를 선별하였다. 이들 선별된 plasmid는 Bacillus sp. SSA3의 expression vector로 사용시 각 재조합 plasmid 중에 삽입된 promoter의 염에 대한 영향의 정도를확인하기 위해 10% NaCl이 첨가된 LB medium상에서 배양하였을 때, 이들 중 Bacillus sp. SSA3의 4 clones은 융합 CAT gene의 발현이 강하게 감소되었으나 2 clones은 약하게 저해되었다.

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Constructions of a Transfer Vector Containing the gX Signal Sequence of Pseudorabies Virus and a Recombinant Baculovirus

  • Lee, Hyung-Hoan;Kang, Hyun;Kim, Jung-Woo;Hong, Seung-Kuk;Kang, Bong-Joo;Song, Jae-Young
    • Journal of Microbiology and Biotechnology
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    • 제9권5호
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    • pp.541-547
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    • 1999
  • Constructions of a transfer vector and a recombinant baculovirus using the thymidine kinase gene of the Herpes simplex virus type 1 strain F (HSV -1) were carried out. Newly cloned transfer vector, pHcgXIIIB, was constructed by insertion of the glycoprotein gX gene signal peptide sequence of Pseudorabies virus into the baculovirus vector pHcEV-IV. The gX sequence was inserted just downstream from the promoter for the polyhedrin gene of the Hyphantria cunea nuclear polyhedrosis virus (HcNPV). HSV-1 thymidine kinase(tk) gene (1.131 kb) was used as a candidate gene for transferring into the baculovirus expression system. The tk gene was inserted into a BamHI site downstream from the gX sequence-promoter for the polyhedrin gene in the pHcgXIIIB transfer vector and was transferred into the infectious lacZ-HcNPV expression vector. Recombinant virus was isolated and was named gX-TK-HcNPV. The recombinant virus produced a 45 kDa gX-TK fusion protein in Spodoptera frugiperda cells, which was confirmed by Western blot analysis. Microscopic examination of gX-TK-HcNPV-infected cells revealed normal multiplication. Fluorescent antibody staining indicated that the gX-TK fusion protein was present in the cytoplasm. These results indicated that the transfer vector successfully transferred the gX-tk gene into the baculovirus expression system.

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기초지반의 지지력보강공법에 관한 연구 (Studies on the Development of Bearing Capacity Reinforcement for the Foundation of Soil)

  • 유동환;최예환;유연택
    • 한국농공학회지
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    • 제30권1호
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    • pp.38-49
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    • 1988
  • This paper presented as follows results of laboratory model tests with various shaped footings on soil bed reinforced with the strips on the base of behaviour of soil structure according to the loads and triaxial test results reinforced with geotextiles. Their parameters studied were the effects on the bearing capacity of a footing of the first layer of reinforcement, horizontal and vertical spacing of layers, number of layers, tensile strength of reinforcement and iclination load to the vertical 1.Depending on the strip arrangement, ultimate bearing capacity values could be more improved than urreinforced soil and the failure of soil was that the soil structure was transfered from the macrospace to microspase and its arrangement, from edge to edge to face to face. 2.The reinforcement was produced the reinforcing effects due to controlling the value of factor of one and permeable reinforcement was never a barrier of drainage condition. 3.Strength ratio was decreased as a linear shape according to increment of saturation degree of soil used even though at the lower strength ratio, the value of M-factor was rot influenced on the strength ratio but impermeable reinforcement decreased the strength of bearing capacity. 4.Ultimate bearing capacity under the plane-strain condition was appeared a little larger than triaxial or the other theoretical formulars and the circular footing more effective. 5.The maximum reinforcing effects were obtained at U I B=o.5, B / B=3 and N=3, when over that limit only acting as a anchor, and same strength of fabric appeared larger reinforcing effects compared to the thinner one. 6.As the LDR increased, more and more BCR occurred and there was appeared a block action below Z / B=O.5, but over the value, decrement of BCR was shown linear relation, and no effects above one. 7.The coefficient of the inclination was shown of minimum at the three layers of fabrics, but the value of H / B related to the ultimate load was decreased as increment of inclination degree, even though over the value of 4.5 there wasn't expected to the reinforcing effects As a consequence of the effects on load inclination, the degree of inclination of 15 per cent was decreased the bearing capacity of 70 per cent but irnproved the effects of 45 per cent through the insertion of geotextile.

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A Yeast MRE3/REC114 Gene is Essential for Normal Cell Growth and Meiotic Recombination

  • Leem, Sun-Hee
    • Journal of Microbiology
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    • 제37권4호
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    • pp.248-255
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    • 1999
  • We have analyzed the MRE3/REC114 gene of Saccharomyces cerevisiae, previously detected in isolation of mutants defective in meiotic recombination. We cloned the MRE3/REC114 gene by complementation of the meiotic recombination defect and it has been mapped to chormosome XIII. The DNA sequence analysis revealed that the MRE3 gene is identical to the REC114 gene. The upstream region of the MRE3/REC114 gene contains a T_4C site, a URS (upstream repression sequence) and a TR (T-rich) box-like sequence, which reside upstream of many meiotic genes. Coincidentally, northern blot analysis indicated that the three sizes of MRE3/REC114 transcripts, 3.4, 1.4 and 1.2 kb, are induced in meiosis. A less abundant transcript of 1.4 kb is detected in both mitotic and meiotic cells, suggesting that it is needed in mitosis as well as meiosis. To examine the role of the MRE3/REC114 gene, we constructed mre3 disruption mutants. Strains carrying an insertion or null deletion of the MRE3/REC114 gene showed slow growth in nutrient medium and the doubling time of these cells increased approximately by 2-fond compared to the wild-type strain. Moreover, the deletion mutant (${\delta}$mre3) displayed no meiotically induced recombination and no viable spores. The mre3/rec114 spore lethality can be suppressed by spo13, a mutation that causes cells to bypass reductional division. The double-stranded breaks (DSBs) which are involved in initiation of meiotic recombination were not detected in the analysis of meiotic chromosomal DNA from the mre3/rec114 disruptant. From these results we suggest that the MRE3/REC114 gene product is essential in normal growth and in early meiotic stages involved in meiotic recombination.

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Optimization of Agrobacterium tumefaciens-Mediated Transformation of Xylaria grammica EL000614, an Endolichenic Fungus Producing Grammicin

  • Jeong, Min-Hye;Kim, Jung A.;Kang, Seogchan;Choi, Eu Ddeum;Kim, Youngmin;Lee, Yerim;Jeon, Mi Jin;Yu, Nan Hee;Park, Ae Ran;Kim, Jin-Cheol;Kim, Soonok;Park, Sook-Young
    • Mycobiology
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    • 제49권5호
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    • pp.491-497
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    • 2021
  • An endolichenic fungus Xylaria grammica EL000614 produces grammicin, a potent nematicidal pyrone derivative that can serve as a new control option for root-knot nematodes. We optimized an Agrobacterium tumefaciens-mediated transformation (ATMT) protocol for X. grammica to support genetic studies. Transformants were successfully generated after co-cultivation of homogenized young mycelia of X. grammica with A. tumefaciens strain AGL-1 carrying a binary vector that contains the bacterial hygromycin B phosphotransferase (hph) gene and the eGFP gene in T-DNA. The resulting transformants were mitotically stable, and PCR analysis showed the integratin of both genes in the genome of transformants. Expression of eGFP was confirmed via fluorescence microscopy. Southern analysis showed that 131 (78.9%) out of 166 transformants contained a single T-DNA insertion. Crucial factors for producing predominantly single T-DNA transformants include 48 h of co-cultivation, pretreatment of A. tumefaciens cells with acetosyringone before co-cultivation, and using freshly prepared mycelia. The established ATMT protocol offers an efficient tool for random insertional mutagenesis and gene transfer in studying the biology and ecology of X. grammica.

접힘홈이 형성된 띠형 섬유보강재를 사용한 패널식 보강토옹벽의 현장계측 연구 (Field Monitoring of Panel-type Reinforced Earth Walls Using Geosynthetic Strip Reinforcement with Folding Grooves)

  • 이광우;조삼덕
    • 한국지반신소재학회논문집
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    • 제17권4호
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    • pp.179-188
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    • 2018
  • 본 연구에서는 새로운 형태의 패널식 보강토옹벽에 대한 현장 적용성 및 구조 안정성을 평가하기 위하여 두 개소 현장에서 현장계측을 수행하였다. 새로운 형태의 패널식 보강토옹벽은 접힘홈이 형성된 띠형 섬유보강재를 패널식 전면벽체에 매립된 C형 삽입구를 통해 직접 연결함으로써 보다 일체화된 구조를 형성시킨 보강토옹벽이다. 현장계측에서는 보강토옹벽의 시공 중 및 완료 후에 발생하는 전면벽체의 수평변위와 띠형 섬유보강재의 인장변형, 전면벽체에 작용하는 수평토압 등을 측정하고 분석하였다. 분석 결과, 보강토옹벽 시공 완료 후에 전면벽체에 작용하는 최대 수평토압은 Rankine 토압의 2/3 이하 수준이고, 전면벽체에 발생된 최대 수평변위는 보강토옹벽 높이의 0.5% 이하 수준이며, 보강재에 유발된 최대 인장변형률은 1.0% 이하로 나타나 두 현장의 보강토옹벽이 모두 안정한 상태를 유지하고 있음을 알 수 있었다.

Comparative mitogenomics of Pleurotus ostreatus Gonji7ho and its cap color mutant

  • Oh, Min-Ji;Na, Kyeong Sook;Jung, Hwa Jin;Lee, Young Kuk;Ryu, Jae-San
    • 한국버섯학회지
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    • 제20권2호
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    • pp.43-49
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    • 2022
  • Pleurotus ostreatus is a globally cultivated mushroom crop. Cap color is a quality factor in P. ostreatus. However, cap color can spontaneously mutate, degrading the quality of the mushroom on the market. Early detection and removal of mutant strains is the best way to maintain the commercial value of the crop. To detect the cap color mutant Gonji7ho, molecular markers were developed based on insertion/deletions (InDels) derived from the comparison of mitogenomes of Gonji7ho and Gonji7hoM mushrooms. Sequencing, assembly, and comparative analysis of the two mitogenomes revealed genome sizes of 73,212 bp and 72,576 bp with 61 and 57 genes or open reading frames (ORFs) in P. ostreatus Gonji7ho and Gonji7hoM, respectively. Fourteen core protein-encoding genes, two rRNA, and 24 tRNA with some OFRs were predicted. Of the 61 genes or OFRs in the wild type, dpo, rpo, and two orf139 were missing (or remnant) in the mutant strain. Molecular markers were developed based on the sequence variations (InDels) between the two mitogenomes. Six polymorphic molecular markers could detect the mutated mitochondria by PCR. These results provide basic knowledge of the mitogenomes of wild-type and mutant P. ostreatus, and can be applied to discriminate mutated mitochondria.

Genomic Characterization and Safety Assessment of Bifidobacterium breve BS2-PB3 as Functional Food

  • Kristin Talia Marbun;Marcelia Sugata;Jonathan Suciono Purnomo;Dikson;Samuel Owen Mudana;Tan Tjie Jan;Juandy Jo
    • Journal of Microbiology and Biotechnology
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    • 제34권4호
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    • pp.871-879
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    • 2024
  • Our group had isolated Bifidobacterium breve strain BS2-PB3 from human breast milk. In this study, we sequenced the whole genome of B. breve BS2-PB3, and with a focus on its safety profile, various probiotic characteristics (presence of antibiotic resistance genes, virulence factors, and mobile elements) were then determined through bioinformatic analyses. The antibiotic resistance profile of B. breve BS2-PB3 was also evaluated. The whole genome of B. breve BS2-PB3 consisted of 2,268,931 base pairs with a G-C content of 58.89% and 2,108 coding regions. The average nucleotide identity and whole-genome phylogenetic analyses supported the classification of B. breve BS2-PB3. According to our in silico assessment, B. breve BS2-PB3 possesses antioxidant and immunomodulation properties in addition to various genes related to the probiotic properties of heat, cold, and acid stress, bile tolerance, and adhesion. Antibiotic susceptibility was evaluated using the Kirby-Bauer disk-diffusion test, in which the minimum inhibitory concentrations for selected antibiotics were subsequently tested using the Epsilometer test. B. breve BS2-PB3 only exhibited selected resistance phenotypes, i.e., to mupirocin (minimum inhibitory concentration/MIC >1,024 ㎍/ml), sulfamethoxazole (MIC>1,024 ㎍/ml), and oxacillin (MIC >3 ㎍/ml). The resistance genes against those antibiotics, i.e., ileS, mupB, sul4, mecC and ramA, were detected within its genome as well. While no virulence factor was detected, four insertion sequences were identified within the genome but were located away from the identified antibiotic resistance genes. In conclusion, B. breve BS2-PB3 demonstrated a sufficient safety profile, making it a promising candidate for further development as a potential functional food.

Comparative Genomic and Genetic Functional Analysis of Industrial L-Leucine- and L-Valine-Producing Corynebacterium glutamicum Strains

  • Ma, Yuechao;Chen, Qixin;Cui, Yi;Du, Lihong;Shi, Tuo;Xu, Qingyang;Ma, Qian;Xie, Xixian;Chen, Ning
    • Journal of Microbiology and Biotechnology
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    • 제28권11호
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    • pp.1916-1927
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    • 2018
  • Corynebacterium glutamicum is an excellent platform for the production of amino acids, and is widely used in the fermentation industry. Most industrial strains are traditionally obtained by repeated processes of random mutation and selection, but the genotype of these strains is often unclear owing to the absence of genomic information. As such, it is difficult to improve the growth and amino acid production of these strains via metabolic engineering. In this study, we generated a complete genome map of an industrial L-valine-producing strain, C. glutamicum XV. In order to establish the relationship between genotypes and physiological characteristics, a comparative genomic analysis was performed to explore the core genome, structural variations, and gene mutations referring to an industrial L-leucine-producing strain, C. glutamicum CP, and the widely used C. glutamicum ATCC 13032. The results indicate that a 36,349 bp repeat sequence in the CP genome contained an additional copy each of lrp and brnFE genes, which benefited the export of L-leucine. However, in XV, the kgd and panB genes were disrupted by nucleotide insertion, which increase the availability of precursors to synthesize L-valine. Moreover, the specific amino acid substitutions in key enzymes increased their activities. Additionally, a novel strategy is proposed to remodel central carbon metabolism and reduce pyruvate consumption without having a negative impact on cell growth by introducing the CP-derived mutant $H^+$/citrate symporter. These results further our understanding regarding the metabolic networks in these strains and help to elucidate the influence of different genotypes on these processes.