• Title/Summary/Keyword: $[U-^{14}C]\

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Construction of a Novel Shuttle Vector for Tetragenococcus species based on a Cryptic Plasmid from Tetragenococcus halophilus

  • Min Jae Kim;Tae Jin Kim;Yun Ji Kang;Ji Yeon Yoo;Jeong Hwan Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.211-218
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    • 2023
  • A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Emr), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.

$\beta$-Mercaptoethanol이 돼지 체외수정란의 생산에 미치는 영향

  • 한만희;구덕본;이경광;박창식;서길웅;정영채;이규승
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.81-81
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    • 2002
  • $\beta$-Mercaptoethanol($\beta$-ME)은 일반적으로 황화합물(thiol compounds)의 일종으로, 배양액 중에서 이황화결합(disulfide bonds)을 분해하여 일정한 물질의 산화.환원반응에 관여하며, 특히 cysteine이 cystine으로 산화되는 것을 차단함으로서 cysteine의 이용능력을 증대시키고, GSH의 합성을 촉진 및 증대시키는 것으로 알려져 있고, 각종 활성산소로부터 세포를 보호하는 역할을 수행하는 것으로 보고되었다. 특히, 돼지수정란의 체외배양체계에 유의적인 영향을 미치는 것으로 보고되었다(Abebydeera 등, Theriogenol., 50:747-756, 1998). 따라서 본 실험에서는 돼지난포란의 체외성숙시 $\beta$-ME의 첨가배양이 체외수정과 배발달에 미치는 영향에 관하여 조사하였다. 돼지난포란을 10% PFF, 0.1mg/ml cysteine, 10IU/m1 PMSG, 10IU/m1 hCG 및 10ng/m1 EGF가 첨가된 NCSU23 배양액에 $\beta$-ME를 각각 0, 25, 50 및 100uM을 처리하여 22시간 동안 배양을 실시하고, 성선자극호르몬이 배제된 배양액에서 추가로 22시간을 배양하여 체외성숙을 유도하였다. 체외성숙이 유기된 난자는 난구세포를 제거하고, 2.5mM caffeine과 0.1% BSA가 첨가된 mTBM배양액에 정자를 1.25 $\times$ $10^{5}$cells/ml의 농도로 5-6시간 동안 공동배양을 실시하여 체외수정을 유도하였다. 체외수정후 일부의 수정란은 12시간에 난자 급속 염색방법으로 염색하여 다정자침입률 및 자.웅전핵형성률 등을 확인하였다. 그리고 나머지1-세포기의 수정란은 0.4mg/ml BSA가 함유된 NCSU23 배양액에 30 embryos/50ul 소적으로하여 38.8$^{\circ}C$, 5% $CO_2$의 탄산가스 배양기에서 각각 7일간 배양을 실시하였다. 조사된 결과는 SAS/STAT를 이용하여 통계분석을 실시하였다. 체외수정 12시간 후에 난자 급속 염색법으로 염색을 실시한 결과, 모든 처리구에서 핵성숙률(76.4~95.2%), 정자침투율(51.1~66.9%), 웅성전핵형성률(95.2~100%), 다정자침입률(18.2~25.6%) 및 평균침입정자수(1.2~l.4개)에서 유의적인 차이가 인정되지 않았다. 체외배양 48시간 난할률을 조사한 결과, 처리구별 차이(53.9~67.9%)는 인정되지 않았으나, 배양 7일째 배반포형성률은 각각 14.5, 25.4, 17.3 및 12.4%로서 25uM의 $\beta$-ME처리구가 유의적(P<0.05)으로 높은 배발달률을 나타내었고, 총세포수에 있어서는 대조구와 처리구간 유의적인 차이가 인정되지 않았다. 따라서 돼지 난포란을 성숙배양할 때, 25uM $\beta$-ME를 첨가배양하는 것이 양질의 돼지체외수정란을 생산하는 하나의 방법으로 조사되었다.다.

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Relationship between Ischemia-Modified Albumin and the Healing Period of Amputation Wounds in Patients with Diabetes Mellitus Following Non-traumatic Below-Knee Amputation (당뇨족으로 인한 비외상성 하퇴부 절단 환자에서 시행한 절단부 창상의 치유 기간과 Ischemia-Modified Albumin과의 관계)

  • Si Young Heo;Myoung Jin Lee;Hyeon jun Kim;Sung Bin Byun
    • Journal of Korean Foot and Ankle Society
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    • v.27 no.2
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    • pp.49-54
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    • 2023
  • Purpose: The present study examined the effectiveness of the preoperative ischemia-modified albumin (IMA) levels in predicting the healing period of amputation wounds in patients with diabetes mellitus following a non-traumatic below-knee amputation (BKA). Materials and Methods: This study enrolled 41 diabetic foot ulcer patients who underwent BKA at the authors' hospital diabetic foot center from April 2016 to April 2022. Among the 41 patients, 29 (70.7%) were male and 12 (29.3%) were female. Their mean age was 64.54±11.38 years (41~81 years). The mean follow-up period was 19.48±5.56 weeks (14~48 weeks) after BKA. The patients were divided into two groups (high IMA group and normal IMA group), which evaluated the healing period, wound dehiscence, and revision operation rate using a Fisher's exact test and Mann-Whitney U test. Three orthopedic surgeons performed stump wound evaluation, and they were evaluated as healing when all sutures were fused without oozing. Results: Thirty patients (73.2%) (group A) showed a high level of IMA (median: 91.2 U/mL), and 11 (26.8%) patients (group B) showed a normal range of IMA (median: 82.7 U/mL). In group A, the median period for wound healing took 1.4 weeks longer, which was significant (p=0.001). No statistical relationship was observed between wound dehiscence, revision operation rate, and IMA value. There was no correlation between the other risk factors (estimated glomerular filtration rate, HbA1c) and the wound healing period. Conclusion: Although there was a limitation in using IMA as the sole factor to predict the healing period of amputation wounds in patients after BKA, this study revealed a significant positive correlation between IMA and the period of stump healing after BKA. Therefore, the preoperative IMA levels may help predict the period of stump healing after BKA.

Corium melt researches at VESTA test facility

  • Kim, Hwan Yeol;An, Sang Mo;Jung, Jaehoon;Ha, Kwang Soon;Song, Jin Ho
    • Nuclear Engineering and Technology
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    • v.49 no.7
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    • pp.1547-1554
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    • 2017
  • VESTA (Verification of Ex-vessel corium STAbilization) and VESTA-S (-small) test facilities were constructed at the Korea Atomic Energy Research Institute in 2010 to perform various corium melt experiments. Since then, several tests have been performed for the verification of an ex-vessel core catcher design for the EU-APR1400. Ablation tests of an impinging $ZrO_2$ melt jet on a sacrificial material were performed to investigate the ablation characteristics. $ZrO_2$ melt in an amount of 65-70 kg was discharged onto a sacrificial material through a well-designed nozzle, after which the ablation depths were measured. Interaction tests between the metallic melt and sacrificial material were performed to investigate the interaction kinetics of the sacrificial material. Two types of melt were used: one is a metallic corium melt with Fe 46%, U 31%, Zr 16%, and Cr 7% (maximum possible content of U and Zr for C-40), and the other is a stainless steel (SUS304) melt. Metallic melt in an amount of 1.5-2.0 kg was delivered onto the sacrificial material, and the ablation depths were measured. Penetration tube failure tests were performed for an APR1400 equipped with 61 in-core instrumentation penetration nozzles and extended tubes at the reactor lower vessel. $ZrO_2$ melt was generated in a melting crucible and delivered down into an interaction crucible where the test specimen is installed. To evaluate the tube ejection mechanism, temperature distributions of the reactor bottom head and in-core instrumentation penetration were measured by a series of thermocouples embedded along the specimen. In addition, lower vessel failure tests for the Fukushima Daiichi nuclear power plant are being performed. As a first step, the configuration of the molten core in the plant was investigated by a melting and solidification experiment. Approximately 5 kg of a mixture, whose composition in terms of weight is $UO_2$ 60%, Zr 10%, $ZrO_2$ 15%, SUS304 14%, and $B_4C$ 1%, was melted in a cold crucible using an induction heating technique.

버섯의 항암효과 -영자 균사체 다당류의 항종양 활성-

  • 이준우
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 1997.06b
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    • pp.13-14
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    • 1997
  • 영지의 약리효과는 triterpenoid계의 저분자와 polysaccharide인 고분자로 대별할 수 있다. 저분자 물질은 항염증작용, 항알러지작용, 간암세포 성장 억제작용, 혈압 강하작용, 혈소판 응집저해작용 등이 있으며, 고분자 물질은 혈압강하작용, 정혈작용, 고지혈증 개선작용, 혈당 강하작용, 면역 증강작용 및 항종양작용 등이 있는 것으로 알려졌다. 현재까지의 영지에 관한 연구는 주로 자실체에 대한 연구가 주류를 이루고 있으며 이들의 산지, 환경조건, 채집시기 및 실험자들에 따라 약리 효과 등에 많은 차이가 있다. 이들에 대한 효과가 영지의 대표적 약리효과라고 언급하기엔 부족한 점이 많으므로 약리효과가 우수하고 일관성이 있는 영지 유래 다당류를 얻기 위한 방법으로서는 균사체의 대량배양법 및 분리방법이 확립 되어야만 한다. 본 연구에서는 새로운 항암활성 다당류의 개발을 위해 국내 자생 영지를 채집하여 검색 및 계통적 분류를 행하였으며 분리된 균주의 최적 액체배양 조건, 다당류 분리 조건, 다당류의 물리화학적 특성 및 약리활성 등에 관한 연구를 실시하였다. 그 중 약리활성이 우수한 G- lucidum IY009 균주를 선발 하였다. 구조적 특징과 종양 면역 활성과의 관계 규명을 위해, G- lucidum IY009 배양균사체로부터 다당류를 T-AS, U-AS, M-AS, S 및 H로 분획 하였다. 추출조건에 따른 다당류 GLG의 수율은 열수 추출 분획이 2.98g/l로 가장 높았으며, U-AS및 T-AS에서 각각 2.32g/l, 2.07g/l이었다. GLG의 특성을 조사하기 위하여 수용성과 비수용성으로 분리한 결과 수용성 분획은 비수용성 분획보다 높은 당 함량을 나타냈으며, T-AS는 70.3%의 당과 7.8%의 단백질로 구성 되었다. GLG 대부분의 분획들은 60~93%의 glucose로 구성된 다당류 이었으며, 주로 $\beta$-glucose로 구성된 다당류 이었다. 아미노산은 Asp 및 Glu의 산성 아미노산과 Ala, Leu 등의 함량이 높게 나타났으며, 비알칼리 추출물에서 Ser과 Thr의 함량이 높게 나타났다. 다당류 T-AS는 평균 분자량이 2,000 kD와 12kD에서 주 peak를 나타냈으며, 수용성 분획의 평균 분자량은 12kD이고 비수용성 분획은 36~2,000 kD의 평균 분자량 분포를 갖는 것으로 나타났다. IR과 NMR 분석 결과 890 cm-1에서 흡수 peak를 나타내어 $\beta$-(1,3)0glucan과 $\beta$-(1,6)-glucan의 구조를 갖는 다당류로 확인 되었다. T-AS 분획은 C:H:O:N의 함량비가 38.9:5.7:49.6:1.84%이며, 이 물질의 융점은 163 $^{\circ}C$로 연한 갈색을 나타낸다. 분리된 GLG의 항암활성 기전 규명을 위해, in vivo 항암실험, 항보체 활성능, 항체 생성능, serum protein 분비능, 대식세포의 탐식능과 활성능 및 세포간 물질 분비 등의 상관관계를 조사하였다. 다당류 GLG 분획물들 가운데 항보체의 활성이 높았던 분획은 sarcome 180에 대한 항암 활성이 높게 나타났다. 다당류 T-AS의 보체 활성화 기작은 classical과 alternative complement pathway의 양 경로를 통해 활성화 되었다. T-AS 분획은 mouse내의 특정 혈청단백을 증가시켰으며, 항체 생성능의 증가가 관찰되어 effect T 세포의 활성화가 나타나고 있음을 알 수 있었다. T-AS는 생체내 투여시에 대식세포의 탐식능이 증진되었으며, 대식세포 기능 저해제에 의한 대식세포의 기능 저해 현상이 회복되었다. 이와 같은 결과들로부터, T-AS의 항암 활성은 활성화된 보체 성분 및 당 수용체들이 존재하는 대식세포의 개입을 시사한다.

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Changes in Physicochemical Properties of Actinidia arguta (Siebold & Zucc.) Planch. ex Miq. by Blanching, Drying, and Fermentation (다래순의 데침, 건조 및 발효 조건에 따른 이화학적 특성 변화)

  • Jeong, Ji-Suk;Kim, Yong-Joo;Park, No-Jin;Go, Geun-Bae;Son, Byeong-Gil
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.3
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    • pp.425-433
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    • 2015
  • This study investigated changes in physicochemical characteristics by drying and fermentation in order to utilize Actinidia arguta. Moisture content of A. arguta was 85.81%. Major sugar and organic acids were sucrose, succinic acid, and citric acid. A. arguta contained 19 kinds of amino acids, including 8 kinds of essential amino acids such as valine, methionine, isoleucine, leucine, threonine, phenylalanine, tryptophan, and lysine. For total organic acids, sample fermented for 17 h at $50^{\circ}C$ under a relative humidity of 80% showed 29,026.53 mg/100 g total organic acids. For total free sugars, cold-dried sample showed the highest level at 6,560.86 mg/100 g, which decreased to 2,386.73 mg/100 g after blanching. For the ratio of essential amino acids, freeze-dried sample showed a content of 11.66%, which increased 4-fold up to 40.71~55.50% with fermentation. Both GABA and vitamin U were highest after 17 h of fermentation (110.29 mg and 6.78 mg/100 g fresh weight, respectively). A. arguta contains a variety of free amino acids that increase in amount after fermentation and thus is expected to be developed as a functional food and substitute tea.

Lactobacillus casei Secreting ${\alpha}$-MSH Induces the Therapeutic Effect on DSS-Induced Acute Colitis in Balb/c Mice

  • Yoon, Sun-Woo;Lee, Chul-Ho;Kim, Jeong-Yoon;Kim, Jie-Youn;Sung, Moon-Hee;Poo, Har-Young
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1975-1983
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    • 2008
  • The neuropeptide ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) has anti-inflammatory property by down regulating the expressions of proinflammatory cytokines. Because ${\alpha}$-MSH elicits the anti-inflammatory effect in various inflammatory disease models, we examined the therapeutic effect of oral administration of recombinant Lactobacillus casei, which secretes ${\alpha}$-MSH (L. casei-${\alpha}$-MSH), on dextran sulfate sodium (DSS)-induced colitis in Balb/c mice. Thus, we constructed the ${\alpha}$-MSH-secreting Lactobacillus casei by the basic plasmid, pLUAT-ss, which was composed of a PldhUTLS promoter and ${\alpha}$-amylase signal sequence from Streptococcus bovis strain. Acute colitis was induced by oral administration of 5% DSS in drinking water for 7 days. To investigate the effect of L. casei-${\alpha}$-MSH on the colitis, L. casei or L. casei-${\alpha}$-MSH was orally administered for 7 days and their effects on body weight, mortality rate, cytokine production, and tissue myeloperoxidase (MPO) activity were observed. Administration of L. casei-${\alpha}$-MSH reduced the symptom of acute colitis as assessed by body weight loss (DSS alone: $14.45{\pm}0.2\;g$; L. casei-${\alpha}$-MSH: $18.2{\pm}0.12\;g$), colitis score (DSS alone: $3.6{\pm}0.4$; L. casei-${\alpha}$-MSH: $1.4{\pm}0.6$), MPO activity (DSS alone: $42.7{\pm}4.5\;U/g$; L. casei-${\alpha}$-MSH: $10.25{\pm}0.5\;U/g$), survival rate, and histological damage compared with the DSS alone mice. L. casei-${\alpha}$-MSH-administered entire colon showed reduced in vitro production of proinflammatory cytokines and $NF-{\kappa}B$ activation. The ${\alpha}$-MSH-secreting recombinant L. casei showed significant anti-inflammatory effects in the murine model of acute colitis and suggests a potential therapeutic role for this agent in clinical inflammatory bowel diseases.

Studies on the Mucilage of the Root of Abelmoschus manihot, MEDIC -[Part V] Kinetics of initial viscosity- (황촉규근(黃蜀葵根) 점액(粘液)에 관한 연구(硏究) -[제오보(第五報)] 초기점도변화(初期粘度變化)의 동역학적고찰(動力學的考察)-)

  • On, Doo-Heayn;Im, Zei-Bin;Sohn, Joo-Hwan
    • Applied Biological Chemistry
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    • v.22 no.1
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    • pp.42-50
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    • 1979
  • The viscosity coefficient of mucilage of Abelmoschus manihot, MEDIC root decrease under the influence of temperature and the other various physical and chemical factors. The rate of viscosity change of the mucilage have been measured at $5^{\circ}{\sim}50^{\circ}C$ in aqueous state under the various conditions. The results are as follows: 1. Relationship between rate of viscosity change of mucilage and temperature can be represent as Andrade equation. 2. Their activation energies of viscosity change of Abelmaschus maihot, MEDIC root A, B, C, D, E, F and G observed are 11.9, 12.1, 11.4, 12.1, 11.6, 13.8 and 13.2 Kcal/mole, respectively. And other activation parameters arc evaluated. 3. The activation energy of naturally mucilage are smaller than that of sterilized mucilage.

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Chemical Resistance of Striped Rice Borer, Chilo Suppressalis, and Green Rice Leafhopper, Nephotettix cincticeps (이화명충과 끝동매미충의 약제저항성)

  • Lee Seung Chan;Yoo Jai Ki
    • Korean journal of applied entomology
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    • v.14 no.2 s.23
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    • pp.65-70
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    • 1975
  • The study was planned to detect resistance levels of striped rice borers (Chilo suppressalis Walker) and green rice leafhoppers(Nephotettix cincticeps Uhler) from different localities to organophosphate and carbamate insecticides using topical application. Strains of overwintering striped rice borers were collected from 7 different areas in Gyeonggi Province and they were kept under conditions of about $25^{\circ}C$ for 10 days. Overwintered and reared larvae of striped rice borers were compared for response to insecticides. The strains of green rice leafhopper were collected from Iri, Milyang, Pyongtaek, Echeon and Suweon. Insects were multiplied in the lab. All insecticides tested were organphosphates(MPP, MEP and Diazinon) and carbamate (NAC), which have been used in control of rice insect pests for over past 10 years. The results obtained were as follows; a. With MPP compound, resistant levels of the Joam, Suweon, and Echeon strains of C. suppressalis were from 2 to 4 times as compared with the Yangpyong strain. b. With MEP insecticide, resistant levels of the Kimpo, Pyongtaek, Yongin, Echeon, Suweon, and Joam strains of C. suppressalis were from 2 to 6 times as compared with the Yangpyong strain. c. With Diazinon, resistant levels of the Suweon strain of C. suppressalis were 3 times greater than the Yangpyong strain; but the resistant levels of Kimpo, Pyongtaek, Yongin, and Echeon strains approximated the later. 4. The overwintered larvae of C. suppressalis were more tolerant to the insecticides than the larvae reared in the lab. e. With MEP, resistant levels of the Iri, Milyang, and Pyongtaek strains of N. cincticeps to the Suweon strain showed 13, 8, 7 and4 times, respectively. f. With MPP, resistant levels of the Iri and Pyongtaek strains of N. cincticeps were 7 and 4 times as compared with the Suweon strain, respectively. g. With NAC, resistant levels of the several strains of if. cincticeps showed no difference, and seemed not to have developed resistance yet.

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Termination of mutant T7 RNA polymerases on intrinsic hairpin-independent termination signal (돌연변이 T7 RNA 증합효소의 머리핀 구조가 만들어지지 않는 인자독립형 전사종결 부위에서의 전사종결에 관한 연구)

  • Shin, Ji-Young;Kim, Dong-Hui;Lee, Sang-Soo
    • The Journal of Natural Sciences
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    • v.14 no.1
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    • pp.25-37
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    • 2004
  • T7 RNA polymerase also recognize intrinsic, hairpin-independent termination signal, a conserved 7-base pair sequence (ATCTGTT in the non-template stand) and U-rich sequence downstream of it. These intrinsic, hairpin-independent termination signal were commonly found in PTH and CJ termination sequences. There are two types of mutant T7 RNA polymerases recognizing sensitively(X3, X19, BG8) of insensitively (R173C) the intrinsic termination signals. We determined the T7 transcription activities of these mutants. Compared to wild-type, mutants X4, 19 and BG8 show highly reduced transcription activities (8%, 33%, 34%). On the other hand mutant R173C shows comparable transcription activity of wild-type (112%). Also transcription termination efficiencies at the PTH or CJ termination signals were determined by using mutant RNA polymerases. Temination of mutnats X4, X19 and BG8 are increased compared to wild-type. On the other hadn mutant R173C proceeds through PTH and CJ termination signals.

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