• Title/Summary/Keyword: pT7MT

Search Result 33, Processing Time 0.025 seconds

pT7MT, a Metallothionein 2A-Tagged Novel Prokaryotic Fusion Expression Vector

  • Marikar, Faiz M.M.T.;Fang, Lei;Jiang, Shu-Han;Hua, Zi-Chun
    • Journal of Microbiology and Biotechnology
    • /
    • v.17 no.5
    • /
    • pp.728-732
    • /
    • 2007
  • In the present article, a novel fusion expression vector for Escherichia coli was developed based on the pTORG plasmid, a derivative of pET32a. This vector, named pT7MT(GenBank Accession No DQ504436), carries a T7 promoter and it drives the downstream gene encoding Metallothionein 2A(MT2A). There are in-framed multiple cloning sites(MCS) downstream of the MT2A gene. A target gene can be cloned into the MCS and fused to the C-terminal of the MT2A gene in a compatible open reading frame(ORF) to achieve fusion expression. The metal-binding capability of MT2A allows the purification of fusion proteins by metal chelating affinity chromatography, known as $Ni^{2+}$-affinity chromatography. Using this expression vector, we successfully got the stable and high-yield expression of MT2A-GST and MT2A-Troponin I fusion proteins. These two proteins were easily purified from the supernatant of cell lysates by one-step $Ni^{2+}$-affinity chromatography. The final yields of MT2A-GST and MT2A-Troponin I were 30mg/l and 28mg/l in LB culture, respectively. Taken together, our data suggest that pT7MT can be applied as a useful expression vector for stable and high-yield production of fusion proteins.

Structure Analysis of pmcABCDEFT Gene Cluster for Degradation of Protocatechuate from Comamonas sp. Strain DJ-12 (Comamonas sp. Strain DJ-12로부터 Protocatechuate의 분해에 관여하는 pmcABCDEFT 유전자군의 구조 분석)

  • Kang Cheol-Hee;Lee Sang-Mhan;Lee Kyoung;Lee Dong-Hun;Kim Chi-Kyung
    • Korean Journal of Microbiology
    • /
    • v.41 no.3
    • /
    • pp.195-200
    • /
    • 2005
  • Comamonas sp. strain DJ-12 is a bacterial isolate capable of degrading of 4-chlorobiphenyl (4CB) as a carbon and energy source. The degradation pathway was characterized as being conducted by consecutive reactions of the meta-degradation of 4CB, hydrolytic dechlorination of 4-chlorobenzoate (4CBA), hydroxylation of 4-hydroxybenzoate, and meta-degradation of protocatechuate to product TCA metabolites. The 6.8 kb fragment from the chromosomal DNA of Comamonas sp. strain DJ-12 included the genes encoding for the meta-degradation of PCA; the genes of protocatechuate 4,5-dioxygenase alpha and beta subunits (pmcA and pmcB), 4-carboxy-2-hydroxymuconate-6-semialdehyde dehydrogenase (pmcC), 2-pyrone-4,6-dicarboxylate hydrolase (pmcD), 4-oxalomesaconate (OMA) hydratase(pmcE), 4-oxalocitramalate (OCM) aldolase (pmcF), and transporter gene (pmcT). They were organized in the order of pmcT-pmcE-pmcF-pmcD-pmcA-pmcB-pmcC. The amino acid sequences deduced from the nucleotide sequences of pmcABCDEFT genes from Comamonas sp. strain DJ-12 exhibited 94 to $98\%$ homologies with those of Comamonas testosteroni BR6020 and Pseudomonas ochraceae NGJ1, but only 52 to $74\%$ with homologies Sphingomonas paucimobilis SYK-6, Sphingomonas sp. LB126, and Arthrobacter keyseri 12B.

Effect of RFLP Marker of the Mitochondrial DNA D-Loop Region on Milk Production in Korean Cattle (한우 Mitochondrial DNA D-Loop 영역의 RFLP Marker가 산유량에 미치는 영향)

  • Chung Eui-Ryong;Chung Ku-Young
    • Food Science of Animal Resources
    • /
    • v.25 no.2
    • /
    • pp.218-225
    • /
    • 2005
  • This study was performed to determine sequence variation and RFLP of the mt DNA D-loop region using Southern blot hybridization analysis and to develop mt DNA marker affecting milk production traits in Hanwoo cows. The PCR was used to amplify an 1142 bp fragment within the D-loop region of mt DNA using specific primers. Mt DNA were digested with seven restriction enzymes and hybridized using DIG-labeled D-loop probe. The mt DNA RFLP polymorphisms were observed in the four enzymes, BamHI, RsaI, XbaI and HpaII. Nucleotide substitutions were detected at positions 441 (G/C), 469 (T/C), 503 (C/T), 569 (G/A), 614 (C/A) and 644 (C/T) of the mt DNA D-loop region between two selected lines. Significant relationship between the XbaI RFLP type and breeding value was found(p<0.05). Cows with A type had higher estimated breeding values than those with B type (P<0.05) between high and low milk production lines. Therefore, the RFLP marker of mt DNA could be used as a selection assisted tool for individuals with high milk producing ability in Hanwoo.

Effect of Soil Mulching after Green Manual Crops on Control of Common Scab and Yield Characteristics of Fall Potato (녹비작물재배후 플라스틱필름 멀칭에 따른 가을감자의 더뎅이병 방제 및 수량특성 변화)

  • 송창길;강봉균
    • Korean Journal of Organic Agriculture
    • /
    • v.8 no.3
    • /
    • pp.99-109
    • /
    • 2000
  • This experiment was conducted to investigate the changes of rate of infected common scab and yield characteristics of fall potato(Solanum tuberosum L.) where green manual crops had been previously cultivated, crushed and tilled and P\ulcornerE transparent vinyl film had been mulched and tunneled to solar heating of soil from May 21. 1998 to July 28. 1998. The total yields of green manual crops which had been previousely cultivated were as followed order : pioneer 855F(64.3MT/ha), soybean(25.0MT/ha), red clover and orchardgrass. The average below-ground temperature at the depths of 5, 10 and 20cm were 54, 45 and 44$^{\circ}C$ during the mulching period, respectively The rate of infected area per potato tuber of common scab decreased by solar heating the soils with mulching after the soybean and red clover cultivation as with previous croppings. Plant height, SPAD(soil plant analysis development) reading, fresh weight of stems, and fall potato's tuber yields tended to increase by the cultivation of green manual crops and mulching of P\ulcornerE film. Fall potato's tuber yields were remarkably affected in the plot of soybean, red clover and pioneer 855F+cultivation of fall potato. T-N, K and Ca contents of fall potato(stem and tuber) also tended to increase by the cultivation of green manual crops and mulching of P\ulcornerE film. T-N, K and Ca contents of soil tended to increase after the cultivation of green manual crops and mulching of P\ulcornerE film and then reduced to contents of soil before green manual planting after potato harvesting.

  • PDF

Role of Chemical Exchange Saturation Transfer and Magnetization Transfer MRI in Detecting Metabolic and Structural Changes of Renal Fibrosis in an Animal Model at 3T

  • Anqin Li;Chuou Xu;Ping Liang;Yao Hu;Yaqi Shen;Daoyu Hu;Zhen Li;Ihab R. Kamel
    • Korean Journal of Radiology
    • /
    • v.21 no.5
    • /
    • pp.588-597
    • /
    • 2020
  • Objective: To investigate the value of combined chemical exchange saturation transfer (CEST) and conventional magnetization transfer imaging (MT) in detecting metabolic and structural changes of renal fibrosis in rats with unilateral ureteral obstruction (UUO) at 3T MRI. Materials and Methods: Thirty-five Sprague-Dawley rats underwent UUO surgery (n = 25) or sham surgery (n = 10). The obstructed and contralateral kidneys were evaluated on days 1, 3, 5, and 7 after surgery. After CEST and MT examinations, 18F-labeled fluoro-2-deoxyglucose positron emission tomography was performed to quantify glucose metabolism. Fibrosis was measured by histology and western blots. Correlations were compared between asymmetrical magnetization transfer ratio at 1.2 ppm (MTRasym(1.2ppm)) derived from CEST and maximum standard uptake value (SUVmax) and between magnetization transfer ratio (MTR) derived from MT and alpha-smooth muscle actin (α-SMA). Results: On days 3 and 7, MTRasym(1.2ppm) and MTR of UUO renal cortex and medulla were significantly different from those of contralateral kidneys (p < 0.05). On day 7, MTRasym(1.2ppm) and MTR of UUO renal cortex and medulla were significantly different from those of sham-operated kidneys (p < 0.05). The MTRasym(1.2ppm) of UUO renal medulla was fairly negatively correlated with SUVmax (r = -0.350, p = 0.021), whereas MTR of UUO renal medulla was strongly negatively correlated with α-SMA (r = -0.744, p < 0.001). Conclusion: CEST and MT could provide metabolic and structural information for comprehensive assessment of renal fibrosis in UUO rats in 3T MRI and may aid in clinical monitoring of renal fibrosis in patients with chronic kidney disease.

Soil Properties of Quercus variabilis Forest on Youngha Valley in Mt. Worak National Park

  • Choi, Hyeon-Jin;Jeon, In-Yeong;Shin, Chang-Hwan;Mun, Hyeong-Tae
    • Journal of Ecology and Environment
    • /
    • v.29 no.5
    • /
    • pp.439-443
    • /
    • 2006
  • Soil properties of Quercus variabilis forest on Youngha valley at Mt. Worak National Park were studied as a part of Korea National Long-Term Ecological Research. Soil sampling was carried out along the 50 cm soil depth with 10cm intervals at every quarter from May 2005 through July 2006. Fresh soil was used for $NH_4{^+}-N,\;NO_3{^-}-N$, and soil water content determination. Remaining soils were air dried in the shade, and then used for determination of soil pH, T-N, T-P and exchangeable cation. Average soil organic matter in top soil was $8.5{\pm}1.2%$ and decreased with soil depth. Bulk density of top soil was $0.82{\pm}0.07g/cm^3 $and increased with soil depth. Soil organic matter and bulk density showed a negative linear correlation ($R^2=0.8464$). Soil pH in top soil and subsoil was similar. T-N, $NH_4{^+}-N,\;NO_3{^-}-N$ and T-P in top soil were $1.9{\pm}0.5mg/g,\;7.3{\pm}1.0mg/kg,\;2.0{\pm}0.4mg/kg\;and\;0.2{\pm}0.05mg/g$, respectively. $K^+,\;Ca^{2+}\;and\;Mg^{2+}$ in top soil were $84.6{\pm}24.4,\;408.8{\pm}137.8\;and\;93.4{\pm}23.0mg/kg$, respectively. They decreased with soil depth. Amounts of organic matter, T-N, $NH_4{^+}-N,\;NO_3{^-}-N$, T-P, $K^+,\;Ca^{2+}\;and\;Mg^{2+}$ in 50 cm soil depth were 250.9, 3.45, 0.025, 0.003, 0.639, 0.181, 0.845 and 0.302 ton $ha^{-1}\;50cm-depth^{-1}$, respectively.

Community of Endophytic Fungi from Alpine Conifers on Mt. Seorak

  • Ju-Kyeong Eo;Ahn-Heum Eom
    • Mycobiology
    • /
    • v.50 no.5
    • /
    • pp.317-325
    • /
    • 2022
  • Endophytic fungi occupy various ecological niches, which reinforces their diversity. As few studies have investigated the endophytic fungi of alpine conifers, we focused on four species of alpine conifers in this study-Abies nephrolepis, Pinus pumila, Taxus cuspidata var. nana, and Thuja koraiensis-and examined them for endophytic fungi. A total of 108 endophytic fungi were isolated. There were four taxa in A. nephrolepis, 12 in P. pumila, 18 in T. cuspidata var. nana, and 17 in T. koraiensis; these were divided into five classes: Agaricomycetes (3.2%), Dothideomycetes (29.0%), Leotiomycetes (15.0%), Sordariomycetes (41.9%), and Orbiliomycetes (1.6%). The most prevalent fungi were Sydowia polyspora (22.7%) and Xylariaceae sp. (22.7%) in P. pumila, Phomopsis juglandina (16.1%) in T. cuspidate var. nana, and Thuja-endophytes sp. 1 (70.0%) in T. koraiensis. However, there was no dominant species growing in A. nephrolepis. Some host plants were analyzed using next-generation sequencing. We obtained 4618 reads for A. nephrolepis and 2268 reads for T. koraiensis. At the genus level, the top three endophytic fungi were Ophiostomataceae_uc (64.6%), Nectriaceae_uc (15.5%), and unclassified organism (18.0%) in A. nephrolepis and Nectriaceae_uc (41.9%), Ophiostomataceae_uc (41.8%), and Magnaporthaceae_uc (9.2%) in T. koraiensis. Our results show that there are different communities of endophytic fungi among different host plants, even if the host plants are in the same region. Such ecological niches are important in terms of the ecological restoration of alpine conifers.

Resolution of the Triacylglycerols Containing Conjugate Trienoic Acids into Their Molecular Species by HPLC in the Reversed-phase and Silver Ion Mode (Reversed-phase 및 $Ag^{+}$-HPLC에 의한 Conjugate Trienoic Acid 함유(含有) Triacylglycerol 분자종(分子種)의 상호분리(相互分離))

  • Kim, Seong-Jin;Woo, Hyo-Kyeng;Joh, Yong-Goe
    • Journal of the Korean Applied Science and Technology
    • /
    • v.18 no.3
    • /
    • pp.197-213
    • /
    • 2001
  • Conjugate trienoic acids (CTA) occurred in triacylglycerols (TGs) of the seed oils of Trichosanthes kirilowii, Momordica charantia and Aleurites fordii, and they were easily converted to their methyl esters in a mixture of sodium methoxide-methanol without any structural destruction. The main fatty acids in triacylglycerol (TG) fraction of the seed oils of Trichosanthes kirilowii are $C_{18:2{\omega}6}$ (32.2 mol %), $C_{18:3{\;}9c.11t,13c}$ (38.0 mol %) and $C_{18:1{\omega}9}$ (11.8 mol %), followed with $C_{16:0}$ (4.8 mol %) and $C_{18:0}$ (3.1 mol %). The TG fraction was resolved into 20 TG molecular species according to the partition number (PN) by reversed-phase (RP)-HPLC. The main TG species were $DT_{c2}$, $MDT_{c}$ and $D_{2}T_{c}$, of which amounts reached 63 mol % of total TG molecular species. The TG sample was fractionated into 11 fractions according to the number of double bond in the molecule by $Ag^{+}-HPLC$ and the species of $DT_{c2}$, $MDT_{c}$ and $D_{2}T_{c}$ were also eluted as main components. The TG species containing CTA showed unusual behaviours in the order of elution by HPLC ; first, TG moleular species of $DT_{c2}$ (D; dienoic acid, $T_{c}$; punicic acid, $T_{ci}$; ${\alpha}-eleostearic$ acid, M ; monoenoic acid, $S_{t}$; stearic acid) was eluted earlier than $Mt_{c2}$, although they have the same PN number of 40, and, secondly, the species of $DT_{ci2}$ with eight double bonds was eluted earlier than that of $D_2T_{ci}$ with seven double bonds. Intact TG of the seed oils of Momordica charantia contained mainly fatty acids such as $C_{18:3{\omega}9c,11t,13t}$ (57.7 mol %), $C_{18:1{\omega}9}$ (17.4 mol %), $C_{18:0}$ (12.3 mol %) and $C_{18:2{\omega}6}$ (10.6 mol %), and was classified into 13 fractions by RP-HPLC. The main TG species were as follows ; $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3\;9c,11t,13t})_{2}$, 39.1 mol %] and $S_{t}T_{ci2}$ [$(C_{18:0})(C_{18:3\;9c,11t,13t})_2$, 33.9 mol %] comprising about 73 mol % of total TG species, accompanied by $DT_{ci2}$ [$(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})_{2}$, 7.3 mol %], $D_{2}T_{ci}$ [$ (C_{18:2{\omega}6})_{2}(C_{18:3\;9c,11t,13t})$, 3.6 mol %] and $MDT_{ci}$ [$(C_{18:1{\omega}9})(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})$, 3.5 mol %]. Simple TG species of $T_{ci3}$ [$(C_{18:3\;9c,11t,13t})_3]$ was present in a small amount of 1.4 mol %, but other simple TG species were not detected. The TG was also resolved into 11 fractions according to the number of double bond by $Ag^{+}-HPLC$, and the species were mainly occupied by $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3\;9c,11t,13t})_{2}$, 39.4 mol %] and $S_tT-{ci2}$ [$(C_{18:0})(C_{18:3\;9c,11t,13t})_{2}$, 35.4 mol %] $DT_{ci2}$ species with eight double bonds was also developed faster than $D_2T_{ci}$ one with seven double bonds as indicated in the analysis of TG of the seed oils of T. kirilowii, and $MT_{ci2}$ species with cis, trans, trans-configurated double bond was eluted earlier than $MT_{c2}$ species with cis, trans, cis-configurated double bond. The main components of fatty acid in total TG fraction isolated from the seed oils of of Aleurites fordii were in the following order ; $C_{18:3\;9c,11t,13t}$ (81.2 mol %)> $C_{18:2{\omega}6}$ (8.5 mol %)> $C_{18:1{\omega}9}$ (5.4 mol %)$. With resolution of the TG by RP-HPLC, eight fractions such as $T_{ci3}$, $Dt_{ci2}$, $D_{2}T_{ci}$, $MT_{ci2}$, $PT_{ci2}$ (P; palmitic acid), $PMT_{ci}$, $PDT_{ci}$ and $S_{t}T_{ci2}$ ($S_{t}$; stearic acid) were isolated, respectively. TG species of $T_{ci3}$ [$(C_{18:3\;9c,11t,13t})_{3}$, 54.2 mol %], $DT_{ci2}$ [$(C_{18:2{\omega}6})(C_{18:3\;9c,11t,13t})_{2}$, 15.0 mol %] and $MT_{ci2}$ [$(C_{18:1{\omega}9})(C_{18:3 9c,11t,13t})_{2}$, 14.8 mol %] were present as main species.

A Study on the Improvement Plan of Gwangju-cheon Water Quality by the Inflow of Mt. Mudeung Valley Water (무등산 계곡수 유입을 통한 광주천 수질 개선 방안에 관한 연구)

  • Ko, Joon-IL;Chung, Seon-Yong
    • Journal of Wetlands Research
    • /
    • v.23 no.3
    • /
    • pp.252-259
    • /
    • 2021
  • Numerous valley waters originating from Mt. Mudeung and flowing into Gwangju Cheon flowed into the confluence-type sewage conduit, the Gwangju Cheon became dry and water quality deteriorated. In this study, a method to create a stream was studied by using the valley water of Mt. Mudeung in the Gwangju cheon that flows into the sewage treatment plant as a water source. Flow and water quality surveys were investigated at four points with meaningful flow quantity. As a result, it showed a flow quantity was 105~2,721 m3/day at each point. And the average water quality was BOD5 0.3~1.6 mg/L. If a stream with a flow quantity of 1,500 m3/day is created during the dry season and then flows into the Namgwang bridge of Gwangju cheon, it is predicted that there will be improvements in BOD 7.3%, COD 6.5%, T-P 5.8%, and T-N 5.2%. In addition, it was determined that the load on the flow quantity of the sewage treatment plant due to the inflow of valley water would be reduced, the cost of sewage treatment would be reduced, and it would be the basis for BGN construction by creating waterside amenity in the city.

Attracting effect of baits used the by-product for swimming crab Portunus trituberculatus pots (부산물을 이용한 꽃게 통발용 미끼의 유인 효과)

  • Chang, Ho-Young;Koo, Jae-Geun;Lee, Keun-Woo;Cho, Bong-Kon;Jeong, Byung-Gon
    • Journal of the Korean Society of Fisheries and Ocean Technology
    • /
    • v.44 no.4
    • /
    • pp.282-293
    • /
    • 2008
  • In order to develop the substitutive materials for natural baits of swimming crab pots, the attracting effects of swimming crab such as the preference of baits which were made of the by-products of marine and stock raising through the water tank experiments and fishing experiments. On the investigation of mean entrapped catch number to the pot by the baits after putting the 4 kinds of baits, mackerel(M), mackerel with grinded mackerel s internals($MM_I$), mackerel with tuna s internals$MM_I$) and makerel with grinded krill(MK) each in one pot by turns, $MM_I$ and MK were entrapped mean 3.9(13.0%) and they were a little more comparing to M, and $MT_I$ is least with mean 2.1(7.0%)(F=12.913, P < 0.05). Otherwise, on the preference investigation of swimming crabs by the baits after putting the 4 kinds of baits in the 4 pots each, M was entrapped mean 3.0(10%), but $MM_I$, $MT_I$ and MK were mean 1.2(4.0%), 1.0(3.3%) and 1.5(5.0%) each and they were only 30-50% of M(F=13.398, P < 0.05). On the preference investigation of swimming crabs by the 5 kinds of baits, mackerel(M), and krill(K), manila clam($M_C$), pig s fat($P_F$) and chicken s head($C_H$) which were used in substitutive baits, M was entrapped mean 3.2(10.7%), but K was about 50% of catch of M with mean 1.6(5.3%), and $M_C$, $P_F$ and $C_H$ were very few with mean 0.1-0.2(0.3-0.7%)(F=89.186, P < 0.05). On the preference investigation of swimming crabs by the pots which were put each the 3 kinds of baits, original krill(K), grinded krill with gluten and soybean oil cake($K_GGS$) and grinded krill with gluten, soybean oil cake and glycine($K_GGSG_L$) in the blue fluorescent hexahedral plastic bait cages(BF), and which were put the mackerel(M) in the non-fluorescent hexahedral red plastic bait cage($RF_N$), it was entrapped mean 3.0(10.0%) in the pot which was put the mackerel in the $RF_N$, and the same level in the pots which were put the K and $K_GGSG_L$ in the BF, but it was mean 2.0(6.7%) in the pots which was put the $K_GGS$ in BF and it was decreased by 30% of catch comparing to $RF_N$(F=3.750, P < 0.05). On the preference investigation of swimming crab by the pots which was put grinded tuna with gluten, soybean oil cake and glycine($T_IGSG_L$) in the blue fluorescent hexahedral plastic bait cage(BF), and which was put mackerel(M) in the nonfluorescent hexahedral red plastic bait cage($RF_N$), it was entrapped mean 3.3(11.0%) in the pot which was put mackerel in $RF_N$, and mean 2.7(9.0%) in the pot which was put $T_IGSG_L$ in BF and it was about 15% less comparing to use bait M(t=1.387, P < 0.05). As a results of fishing experiments, a plan for enhancing catching efficiency of $T_IGSG_L$ will be required because catching efficiency of $T_IGSG_L$, alternative bait, was half of fish catching efficiency of natural bait using mackerel. Fishing experiments were conducted 3 times using reinforced substitutive artificial bait that is reinforced attractive effect of $T_IGSG_L$ and composed of tuna intestine, grinded mackerel, gluten, soybean cake, glycine and alanine($T_IM_GGSG_LA$). Catching efficiency of $T_IM_GGSG_LA$ was about 80% of that of natural bait made of mackerel.