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염기서열 특이적 전사종결에 영향을 주는 T7 파아지 lysozyme의 역할 (Role of T7 phage lysozyme affected sequence-specific transcription termination by T7 RNA polymerase)

  • 김동희;이상수
    • 자연과학논문집
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    • 제14권1호
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    • pp.51-61
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    • 2004
  • 박테리오 파아지 T7 RNA 중합효소는 어떤 전사인자도 관여하지 않는 간단한 시스템으로 파아지 RNA 중합효소와 전사촉진제만의 단백질-DNA 상호작용에 의해 전사가 진행된다. T7 파아지의 숙주 세포의 파괴에 관여하는 T7 파아지 lysozyme은 전사를 억제하고, 전사종결에 영향을 미친다. 따라서 T7 파아지 lysozyme 유전자를 대장균 발현 벡터에 삽입하여 pT7lys를 얻었고, 발현시켜 Ni-NTA column chromatography로 순수 분리하였다. T7 파아지 lysozyme은 SDS-gel에서 단일 밴드로 확인하였으며, amidase 활성 역시 확인하였다. 염기서열 특이적 전사 종결에 미치는 T7 파아지 lysozyme의 역할을 알아보기 위하여, rrnB T1 전사종결 인자 부근에서의 전사연장 복합체 제조에 미치는 T7 파아지 lysozyme의 영향력을 조사하였다. 이 결과 T7 파아지 lysozyme 존재 하에 형성되는 전사연장 복합체는 불안정함을 알 수 있었다.

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박테리오파아지 T7 의 기능에 관한 연구;복제단백질간의 단백질 상호작용 (Funcyional Studies on Gene 2.5 Protein of Bacteriophage T7 : Protein Interactions of Replicative Proteins)

  • 김학준;김영태
    • 생명과학회지
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    • 제6권3호
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    • pp.185-192
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    • 1996
  • 박테리오파지 T7 gene 2.5 단백질은 single-stranded DNA 결합 단백질로 박태리오파지 T7의 DNA복제, 재조합, 및 수선에 필수적으로 요구된다. Gene 2.5 protein은 T7의 DNA 합성과 성장에 필수적인 단백질이다. Gene 2.5 Protein이 중요시 되는 이유는 이 단백질이 T7의 다른 복제 필수단백질인 T7의 다른 복제 필수단백질인 T7 DNA polymerase 와 gene 4 protein(helicase/primase)와 서로 상호작용할 것으로 제안되었기 때문이다. (Kim and Richardson, J. Biol. Chem., 1992;1994). 이 단백질의 단백질 상호작용을 가능하게 하는 domain은 carboxyl-terminal domain일 것으로 여러 실험에서 대두되었기에, 이 domain의 특성을 파악하기 위해 야생형과 변이체 gene 2.5 단백질들을 각각 GST에 융합한후 fusion 단백질을 정제하였다. 정제된 이 융합 단백질들의 carboxyl-terminal domain이 T7 복제 단백질들과 상호작용을 조사하는지를 조사하기 위해 affinity chromatography로 이용하였다. 실험 결과, 아생형 GST-gene 2.5 융합단잭질(GST-2.5 (WT))는 T7 DNA polymerase 와 상호작용을 하였지만. 변이형 융합단백질(GST-2.5$\Delta$21C)는 interaction을 하지 못했다. 이 결과는 carbohyl-terminal domain이 단백질-단백질 상호작용을 하는데 직접적으로 관여하는 것을 증명하였다. 또한,GST2.5(WT)는 gene 4 protein(helicase/primase)와 직접 상호작용을 하나. GST2.5$\Delta$21C는 상호작용을 하지 못하는 것으로 나타났다. 따라서 gene 4 proteins와의 상호작용에도 gene 2.5 protein의 carboxyl-terminal domain이 직접 관여 한다는 것이 증명되었다. 이상의 결과에서 gene 2.5 protein은 박테리오파지 T7 의 유전자 목제 시 단백질-단백질 상호작용에 관혀아며, 특히 gene 2.5 protein의 carboxyl-terminal domain이 이러한 상호작용에 직접적으로 관여하는 domain이라는 것을 알 수가 있었다.

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T7 RNA polymerase 유전자의 담배식물에서의 발현 (T7 RNA Polymerase Is Expressed in Plants in a Nicked but Active Form)

  • ;;박상규
    • Applied Biological Chemistry
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    • 제40권4호
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    • pp.271-276
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    • 1997
  • 박테리오파아지 T7 RNA polymerase 유전자를 식물체내에서 이용할 수 있을지 알아보기 위하여 상처유발인 감자 단백질 분해효소 억제제 유전자의 프로모터에 박테리오파지 T7 RNA polymerase 유전자를 연결시킨 후 담배에 도입시켰다. 형질전환 식물체의 DNA에 대한 Southern hybridization에 의하면 T7 RNA polymerase 유전자가 식물체내에 1-2 copy가 존재하며, Northern hybridization에 의하면 T7 RNA polymerase의 RNA가 상처에 따라 생성되는 것을 확인하였다. 또한 Western hybridization에 의하면 식물체내 T7 RNA polymerase 단백질이 생성되는데 그 크기는 대장균에서 생성되는 단백질 크기와 유사한 80 kDa 이었으며 시험관내에서 전사체에 뉴클레오타이드를 결합시키는 능력이 있음도 확인하였다. 따라서 T7 RNA polymerase 유전자를 이용하여 식물체내에서 원하는 유전자의 발현을 증대시킬 수 있을 것으로 사료된다.

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Efficacy of Cyanobacterial Biofertilizer (CBB) on Leaf Yield and Quality of Mulberry and its Impact on Silkworm Cocoon Characters

  • Dasappa D.M. Ram Rao;Ramaswamy S.N.
    • International Journal of Industrial Entomology and Biomaterials
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    • 제13권1호
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    • pp.15-22
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    • 2006
  • An experiment was conducted to study the efficiency of cyanobacterial biofertilizer (CBB) with chemical (NPK) fertilizer on quantitative and qualitative characters of mulberry variety Kanva-2. Their influences on silkworm growth and cocoon characters were also studied. Ten different CBB and NPK fertilizer treatments were given to 5000 plants of established mulberry garden. Treatments were of four types viz., (i) T1 to T7: single and combination dose of CBB+50% NPK (ii) T8: combination dose of CBB + 25%NPK, (iii) T9: CBB only and (iv) T10: control-l00% NPK. Soil pH decreased and nutrients status increased in CBB (T1- T9) treated plots. Average of ten crops data on quantitative traits revealed that T7 (CBB [N. muscorum (1.0 g), A. variahilis (1.0) and S. millei (1.0 g)] + 50% NPK) was very effective in improving growth parameters. Leaf yield was also found high in treatment T7 (32.12 tons/ha/yr.) followed by T10 (31.17 tons/ha/yr.) and T8 (27.67 tons/ha/yr.). Leaf quality characters were found high in T7 and low in T9. Most of the quality traits in T7 are on par with control no. The results revealed that reduction in the dose of chemical fertilizers in T7 did not affect the leaf yield and leaf quality traits of mulberry. This clearly indicates that the efficiency of CBB (T7) provides nitrogen, increases essential nutrients available in soil, maintain soil pH and supply growth substances required for the improvement of leaf yield and leaf quality of mulberry. Bioassay study also revealed no significant difference in silkworm growth and cocoon characters between treatments T7 and T10. Economics calculated revealed that T7 is highly economical and beneficial over T10 by gaining an amount of Rs. 660/-/acre/crop. Thus, treatment T7 containing N. muscorum (1.0 g), A. variahilis (1.0 g) and S. millei (1.0 g) + 50% NPK fertilizers can be recommended to sericulturists mainly to reduce the use of NPK fertilizers, by saving 50% of its cost and to improve soil fertility conditions, which in turn improves leaf yield and quality of mulberry.

생육단계별 예취 시기가 수수 X 수단그라스 교잡종의 이용회수, 생육특성, 수량 및 조단백질수량에 미치는 영향 (Effect of Cutting Times according to Growth Stage in Sorghum X Sudangrass Hybrid on Frequence of Use, Growth Characteristics, Forage Production and Crude Protein Yield)

  • 전병태;이상무
    • 한국초지조사료학회지
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    • 제25권1호
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    • pp.33-42
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    • 2005
  • A field experiment was conducted to evaluate growth characteristics, forage production and crude protein yield according to cutting time of Soghum ${\times}$ Sudangrass Hybrid, and decide ideal harvesting time for use of soiling and silage. Experiment design was arranged with 7 different treatment T1(150 m), T2(200 cm), T3(boot), T4(heading), T5(milk), T6(dough) and T7(yellow stage), as a randomized block design. The results were as fellows : Cutting times of utilization during the course of a year was 4 times at T1 and T2, 3 times at T3 and T4, and 2 times at T5, T6 and T7. Accumulative plant length was the highest at T2(666cm), but T3 was the lowest as 402 cm. Mean Leaf length was the highest at T5(82.1 m) and lowest at T7(T1.8 m). Mean leaf width was the highest at T2 and lowest at T6. Stem diameter was orderly ranked as T3(10.7 mm)>T1(9.5)>T2, T5(9.3>T6(8.9)>T7(8.6)>T4(8.5). Stem hardness was orderly ranked as $T7(3.2 kg/cm^2$>T5, T6(2.3)>T3, T4(1.5)> T2(0.6)>T7(8.6)>T1(0.5). Mean of leaf number and leaf ratio was the highest at $T3(8.1\%)$ and $T2(45.3\%)$, respectively. The highest yield of fresh and dry matter was obtained at T4 and T6 as 113,246 and 24,249 kg/ha, respectively(P<0.05), and e lowest at T7 and T1 as 82,675 and 13,006 kg/ha, respectively(P<0.05). Crude protein yield was highest at T6(1.456 kg/ha) and lowest at T3 as 1,189 kg/ha. As mentioned above the result T1, T2 and T3 could be recommended as use of soiling, and T5, T6 and T7 as silage.

T7 발현체계에서 chloramphenicol acetyltransferase의 선택적 과잉생산 (Selective overproduction of chloramphenicol acetyltransferase in the T7 expression system)

  • 김한복;강창원
    • 미생물학회지
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    • 제27권4호
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    • pp.317-322
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    • 1989
  • A gene can be selectively overexpressed in E. coli by utilizing the phage T7 RNA polymerase's stringent recognition and active transcription of the T7 promoter. The T7 expression system was constructed such that the T7 RNA polymerase gene is under the control of lacUV5 promoter in one plasmid, and that the target gene, the promoterless chloramphenicol acetyltransferase (CAT) gene with E. coli ribosome binding site is under the control of T7 promoter in the other plasmid. Only the E. coli cells containing both plasmids show high resistance to chloramphenicol. When the copy number of the runaway plasmid containing the polymerase gene was varied by a temperature shift, amounts of the CAT protein synthesized upon induction was correspondingly changed as shown in SDS gel electrophoresis.

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Analyzing the Significance of T1 Slope minus Cervical Lordosis in Patients with Anterior Cervical Discectomy and Fusion Surgery

  • Lee, Ho Jin;You, Soon Tae;Sung, Jae Hoon;Kim, Il Sup;Hong, Jae Taek
    • Journal of Korean Neurosurgical Society
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    • 제64권6호
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    • pp.913-921
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    • 2021
  • Objective : Accurate measurement of T1 slope (a component of T1s minus cervical lordosis [CL]) is often constrained by anatomical limitations. In this situation, efforts should be made to find the exact meaning of T1s-CL and whether there are any alternatives to it. Methods : We enrolled 117 patients who received two-level anterior cervical discectomy and fusion (ACDF). Occipital slope, C2 slope (C2s), C7 slope (C7s), T1, O-C2 angle (O-C2A), C2-7 angle (C2-7A), O-C7 angle (O-C7A), T1s-CL, C7-T1 angle (C7-T1A), and C2-7 sagittal vertical axis were measured. We determined 16° (T1s-CL) as the reference point for dividing subjects into the mismatch group and the balance group, and a comparative analysis was performed. Results : The mean value of C7-T1A was constantly maintained within 2.6° peri-operatively. In addition, C2s and T1s-CL showed the same absolute change (Δ|0.8|°). The mean values of T1s-CL of the mismatch and balance groups were 23.0° and 7.6°, respectively. The five factors with the largest differences between the two groups were as follows : C2s (Δ13.3°), T1s-CL (Δ15.4°), O-C2A (Δ8.7°), C2-7A (Δ14.7°), and segmental angle (Δ7.9°) before surgery. Only four factors showed statistically significant change between the two groups after ACDF : T1s-CL (Δ4.0° vs. Δ0.2°), C2s (Δ3.2° vs. Δ0.7°), O-C2A (Δ2.6° vs. Δ1.3°), C2-7A (Δ6.3° vs. Δ1.3°). A very strong correlation between T1s-CL and C2s was also found (r=|0.88-0.96|). Conclusion : C2s itself may be the essential key to represent T1s-CL. The amounts and directions of change of these two factors (T1s-CL and C2s) were also almost identical. The above phenomenon was re-confirmed once again through the correlation analysis.

두정부 백질 물질을 이용한 수소 자기 공명 분광 분석 (Analysis of $^1H$ MR Spectroscopy of parietal white matter material Phantom)

  • 이재영;임청환;김명수
    • 대한방사선기술학회지:방사선기술과학
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    • 제26권2호
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    • pp.57-61
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    • 2003
  • 두정부 백질 물질 NAA, Choline(Cho), Creatine(Cr), choline chloride, glutamin 등으로 자체 제작한 phantom을 이용하여 고 자장(4.7T)과 중자장(1.5T)에서의 신호강도 대 잡음비, 스펙트럼 분해능을 알아 볼 수 있는 선폭과 $T_2$, 그리고 $T_E$값의 변화에 따른 각 대사물질의 변화별 스펙트럼 등을 구하여 이론적인 정보와 실제적인 정보와의 차이를 알아보고자 한다. 이용된 기기는 Bruker Biospec 4.7T와 1.5T GE SIGNA를 이용하여 MRS의 결과를 얻었다. 관심영역에서 얻은 정보에서 분광 peak의 면적을 구하였다. 통계처리는 Microsoft사의 Excel program내에 있는 통계 package를 이용하였다. 중자장(1.5T)과 고자장(4.7T)에서의 각각의 대사물질의 스펙트럼을 얻어 본 결과 자장의 균일도와 SNR과 $T_2$값의 차이가 이론적인 값보다는 직선성을 보이지 않았다. 1.5T에서의 선폭은 Cho, Cr, NAA 순서로 $5.30{\pm}1.07,\;4.81{\pm}0.14,\;5.49{\pm}0$, 이에 반해 4.7T에서는 $9.14{\pm}0.55,\;8.87{\pm}0.67,\;9.65{\pm}0.56$ Hz 값이 나왔다. 평균 SNR은 NAA의 물질의 경우를 3회 측정하였는데 그 평균치는 63%의 증가를 보였다. $T_E$ 변화로 스펙트럼을 분석했을 때 $T_E=60ms$일 때 가장 SNR이 좋은 값을 보였고 $T_E=135ms$일 때 가장 낮은 값을 나타나 보였다. 두정부 백질 물질 Phantom을 제작하여 자장의 강도 즉 중자장(1.5T)과 고자장(4.7T)에 따라 분석하게 되었는데 고자장의 영역으로 갈수록 분해능, SNR은 좋아져 보이나 자장의 세기가 커질수록 이론적인 수치보다는 상당히 적은 증가를 보이고 있다는 것을 이 연구를 통해 알게 되었다.

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Characterization of a Bacteriocin Produced by Enterococcus sp. T7 Isolated from Humans

  • Moon, Hi-Seong;Jeong, Jong-Jin;Ji, Geun-Eog;Kim, Jong-Sang;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제10권4호
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    • pp.507-513
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    • 2000
  • A bacteriocin-producing organism, Enterococcus sp. T7, was isolated from human fecal samples. Bacteriocin T7, named tentatively as the bacteriocin, was produced by Enterococcus sp. T7 and it inhibited some strains of Lactobacillus. Staphylococcus, Enterococcus, and Streptococcus, but not all the lactococci and gram-negative bacteria tested. Bacteriocin T7 inhibited the growth of Listeria monocytogenes Scott A, but the degree of inhibition was less than those for other sensitive gram-positive vacteria. Bacteriocin T7 in MRS broth started to produce at the middle of the exponential growth phase and the inhibitory activity reached its maximum level during the stationary growth phase. Bacteriocin T7 was stable against heat treatments, pH variations (pH 2-10), and exposure to organic solvents. The molecular weight of bacteriocin T7 was estimated to be 6.500 Da by SDS-PAGe. All these facts, including physico-chemical stabilities, small molecular size, and inhibition of Kisteria monocytogenes, indicate that bacteriocin T7 is likely to be a member of the class IIa bacteriocins.

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Seeing is Believing: Illuminating the Source of In Vivo Interleukin-7

  • Kim, Grace Yoon-Hee;Hong, Chang-Wan;Park, Jung-Hyun
    • IMMUNE NETWORK
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    • 제11권1호
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    • pp.1-10
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    • 2011
  • Interleukin-7 (IL-7) is an essential cytokine for T cells. However, IL-7 is not produced by T cells themselves such that T cells are dependent on extrinsic IL-7. In fact, in the absence of IL-7, T cell development in the thymus as well as survival of naive T cells in the periphery is severely impaired. Furthermore, modulating IL-7 availability in vivo either by genetic means or other experimental approaches determines the size, composition and function of the T cell pool. Consequently, understanding IL-7 expression is critical for understanding T cell immunity. Until most recently, however, the spatiotemporal expression of in vivo IL-7 has remained obscured. Shortage of such information was partly due to scarce expression of IL-7 itself but mainly due to the lack of adequate reagents to monitor IL-7 expression in vivo. This situation dramatically changed with a recent rush of four independent studies that describe the generation and characterization of IL-7 reporter mice, all utilizing bacterial artificial chromosome transgene technology. The emerging consensus of these studies confirmed thymic stromal cells as the major producers of IL-7 but also identified IL-7 reporter activities in various peripheral tissues including skin, intestine and lymph nodes. Strikingly, developmental and environmental cues actively modulated IL-7 reporter activities in vivo suggesting that IL-7 regulation might be a new mechanism of shaping T cell development and homeostasis. Collectively, the availability of these new tools opens up new venues to assess unanswered questions in IL-7 biology in T cells and beyond.