• Title/Summary/Keyword: KB-C2

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Expression of Human Serum Albumin in Milk of Transgenic Mice Using Goat β-casein/Human Serum Albumin Fusion Gene

  • Wu, H.T.;Chou, C.K.;Huang, M.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.6
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    • pp.743-749
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    • 2004
  • The gene encoding human serum albumin (HSA) was cloned from human liver cDNA library by PCR. The HSA cDNA in size of 2,176 bp, including 1,830 bp of open reading frame, was cloned into the plasmid carried with the 5'flanking sequence of goat $\beta$-casein gene (-4,044 to +2,025 bp) to get a tissue specific expression vector in mammary gland named pGB562/HSA (12.5 kb). A 9.6 kb DNA fragment in which the sequence is in order of goat $\beta$-casein gene regulatory sequence, HSA cDNA and SV40 polyadenylation signals was isolated from the pGB562/HSA by SacI and DraIII cutting, and used to microinject into the pronuclei of mouse fertilized eggs to produce transgenic mice. Three transgenic mice (2 female and 1 male) were identified by PCR and dot Southern blot analysis. The copy numbers of integrated transgene were more than 10 copies in line #21 and #26 as well as over 50 copies in line #31 of transgenic mice. HSA protein collected from the milk of lactating transgenic mice was confirmed by immuno-detection of Western and slot blot. The concentrations of HSA in the milk were from 0.05 to 0.4 mg/ml. An obvious antigen and antibody conjugate could be observed in immunohistochemical stain of mammary gland tissue from lactating day 11 of HSA transgenic mice. The transmission of transgene and its expression was recognized according to the results of RT-PCR and sequences analyses of their progeny.

AN EXPERIMENTAL STUDY ON THE CHANGE OF THE RADIOSENSITIVITY OF SEVERAL TUMOR CELL LINES AND PRIMARY CULTURED GINGIVAL FIBROBLAST (수종의 암세포주와 치은섬유아세포에서 방사선의 양과 분할조사에 따른 세포활성도와 독성의 변화에 관한 연구)

  • Lee Sam-Sun;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.27 no.1
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    • pp.107-122
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    • 1997
  • Radiation sensitivity data was generated for two human cancer cell lines(KB, RPMI 2650) and human primary gingival fibroblast was tested three times using a viable cell number counting with a hemocytometer, MTT(3-[4,5-Dimethylthiazol2-yl]-2,5-diphenyl tetrazolium bromide) assay, and LDH(Lactate dehydrogenase) assay. Single irradiation of 2, 4, 6, 10, 15, 20Gy were applied to the tumor cell lines and the primary cultured gingival fibroblast The two fractions of 4Gy and 10Gy were seperated with a 4 hour time interval. The irradiation was done with 241.5cGy/min dose rate using /sup 137/Cs MK cell irradiator at room temperature. The obtained results were as followed : 1. There was significantly different viable cell numbers as the amount of radiation dose on the tested cells were cell number counted with a hemocytometer. In fractions, there were more viable cells remaining. 2. Phase-contrast microscopically, radiation-induced morphologic changes were pronounced on the tumor cells, however, almost no differences on the gingival fibroblast. 3. There was significantly different absorbance at 2Gy on RPMI 2600, 4Gy on KB and GF in MTT assay. In fractions, the absorbance was significantly higher on KB. 4. The level of extracellular LDH activity in the experimental group was significantly higher in the 2-4Gy than the control group. 5. The total level of extracellular and intracellular LDH activity was decreased as increased amounts of radiation dose was applied.

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Isolation and characterization of sodium dodecylbenzenesulfonate(soft type)-degrading bacteria (Sodium dodecylbenzenesulfonate(soft type)를 분해하는 미생물의 분리 및 특성)

  • 전홍기;안영희;백형석
    • Korean Journal of Microbiology
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    • v.27 no.3
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    • pp.279-284
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    • 1989
  • Macroorganisms capable of utilizing sodium dodecylbenzenesulfonate(SDBS, soft type) as a sole carbon source were isolated from nature by using SDBS agar plate technique. Iwolated bacteria were examined primarily for biodegradation ability of SDBS, and followed by testing for resistance to several kinds of metal compounds and antibiotics. Among them(152 strains), one strain showed a excellent SDBS-degrading ability with a resistance to amipicillin and rifampicin was selected. This bacterium was identified as Klebsiella sp. and harbored two plasmids of about 4 and 5 kilobases. SDBS-degrading ability was lost when the plasmids were cured by mitomycin C. It was revealed that the degradation of SDBS was controlled by the plasmid DNA encoding genes. The two plasmids were stably maintained in Escherichia coli C600.

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Study of reaction mechanism in pre-reforming for MCFC (MCFC의 예비 개질 반응 메커니즘 연구)

  • Lee, Woo-Hyung;Park, Yong-Ki
    • Industry Promotion Research
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    • v.3 no.2
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    • pp.1-8
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    • 2018
  • In this study, the reaction mechanism of ethane and the reaction rate equation suitable for hydrocarbon reforming were studied. Through the reaction mechanism analysis, it was confirmed that three reactions (CO2 + H2, C2H6 + H2, C2H6 + H2O) proceed during the reforming reaction of ethane, each reaction rate (CO2+H2($r=3.42{\times}10-5molgcat.-1\;s-1$), C2H6+H2($r=3.18{\times}10-5mol\;gcat.-1s-1$), C2H6+H2O($r=1.84{\times}10-5mol\;gcat.-1s-1$)) was determined. It was confirmed that the C2H6 + H2O reaction was a rate determining step (RDS). And the reaction equation of this reaction can be expressed as r = kS * (KAKBPC2H6PH2O) / (1 + KAPC2H6 + KBPH2O) (KA = 2.052, KB = 6.384, $kS=0.189{\times}10-2$) through the Langmuir-Hinshelwood model. The obtained equation was compared with the derived power rate law without regard to the reaction mechanism and the power rate law was relatively similar fitting in the narrow concentration change region (about 2.5-4% of ethane, about 60-75% of water) It was confirmed that the LH model reaction equation based on the reaction mechanism shows a similar value to the experimental value in the wide concentration change region.

Construction of Various Copy Number Plasmid Vectors and Their Utility for Genome Sequencing

  • Yang, Tae-Jin;Yu, Yeisoo;Frisch, David A.;Lee, Seunghee;Kim, Hye-Ran;Kwon, Soo-Jin;Park, Beom-Suk;Wing, Rod A.
    • Genomics & Informatics
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    • v.2 no.4
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    • pp.174-179
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    • 2004
  • We developed various plasmid cloning vectors that are useful in the construction of genomic and shotgun libraries. Two medium copy vectors, pCUGlblu21 (pCb21) and pAGlblu21 (pAb21), which are resistant to kanamycin ($Km^R$) and chloramphenicol ($Cam^R$), respectively, are useful for cloning DNA inserts ranging from 5kb to 15kb. Two high copy vectors, pCUGlblu31 (pCb31) and pAGlblu31 (pAb31), containing $Km^R$ and $Cam^R$, respectively, are useful for DNA inserts less than 5kb. These vectors are well adapted for large-scale genome sequencing projects by providing choice of copy number and selectable marker. The small vector size is another advantage of these vectors. All vectors contain lacZa including multicloning sites that originated from pBluscriptllsk- for easy cloning and sequencing. Two medium copy vectors contain unique and rare cutting Swal (ATTTAAAT) restriction enzyme sites for easy determination of insert size. We developed two combined vectors, pC21A31 and pC31A21, which are combinations of (pCb21 + pAb31) and (pCb31 + pAb21), respectively. These two vectors provide four choices of vectors such as $Km^R$ and medium, $Cam^R$ and high, $Cam^R$ and medium, and $Km^R$ and high copy vectors by restriction enzyme cutting, dephosphorylation, and gel purification. These vectors were successfully applied to high throughput shotgun sequencing of rice, tomato, and brassica BAC clones. With an example of extremely biased hydro sheared 3 kb shotgun library of a tomato BAC clone, which is originated from cytogenetically defined peri-centromeric region, we suggest the utility of an additional 10 kb library for sequence assembly of the difficult-to-assemble BAC clone.

Petrogenesis and Metamorphism of Charnockite of Eastern Jirisan Area (지리산 동부 지역에 분포하는 차노카이트의 변성작용과 성인에 관한 연구)

  • 김동연;송용선;박계헌
    • The Journal of the Petrological Society of Korea
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    • v.11 no.3_4
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    • pp.138-156
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    • 2002
  • Precambrian metamorphic rocks of southwest Sobaeksan massif consist of mainly granitic gneiss, porphyroblastic gneiss and quartzofeldspathic gneiss. The orthopyroxene-bearing rocks(charnockites) are found in the west of Hadong-Sancheong anorthosite complex. The charnockites are 3km wide, 12km long and divided into massive and foliated types based on their texture. The compositions of charnockites are comparable to granodiorite to adamellite and subalkaline. Variations in major and trace elemental abundances show typical magmatic differentiation trends. The geochemical data plotted on tectonic discrimination diagrams reveal that these charnockites were formed in the active tectonic environment. The massive and folidated charnockites are mainly composed of plagioclase, orthopyroxene, microcline, quartz and disseminated garnet. Camels generally show characteristic zonal textures with decreasing $X_{alm}$(0.74~0.83), $X_{Py}$ (0.07~0.12) and $X_{Mg}$ (0.12~0.08) and increasing $X_{grs}$(0.03~0.15) from core to rim. Metamorphic temperature and pressure of the charnockites estimated from orthopyroxene-garnet-plagioclase-quartz assemblages show wide range of variation of $600~900^{\circ}C$ and 2.5~7.5 kbar respectively. The results of P-T estimates indicate an anticlockwise P-T evolution path.

Pepstatin- Insensitive Carboxyl Proteinase: A Biochemical Marker for Late Lysosomes in Amoeba proteus

  • Hae Kyung Kwon;HyeonJung Kim;Tae In Ahn
    • Animal cells and systems
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    • v.3 no.2
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    • pp.221-228
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    • 1999
  • In order to find a biochemical marker for late Iysosomes, we characterized two cDNAs which were cloned by using a monoclonal antibody (mAb) against Iysosomes in Amoeba proteus as a probe. The two cDNAs, a 1.3-kb cDNA in pBSK-Iys45 and a 1.6-kb cDNA in pBSK-Iys60, were found to encode proteins homologous to pepstatin-insensitive carboxyl proteinases (PICPs). E. coli transformed with pBSK-Iys45 produced two immunopositive polypeptides (45 and 43 kDa) and the cDNA in 1274 bases encoded a 44,733-Da protein (Lys45) of 420 amino acids containing one site for a core oligosaccharide. On the other hand, E. coli transformed with pBSK-Iys60 produced several polypeptides (64, 54, 45, 41, and 37 kDa) reacting with the mAb. The cDNA contained 1629 bases and encoded a 59,231-Da protein (Lys60) of 530 amino acids containing two sites for asparagine-linked core oligosaccharides. These two cDNAs showed identities of 60.3% in nucleotide sequences and 23.6% in amino acid sequences. Lys45 and Lys60 appeared to share XXEFQK as a common antigenic domain. The amino acid sequence of the Lys45 protein showed 17.4% identity and 40.9% similarity to that of PICP from Pseudomonas sp. 101. On the other hand, Lys60 showed a 24.3% identity and 51.9% similarity with human Iysosomal PICP in the amino acid sequence. A putative active center for serine protease, GTS*xxxxxFxG, was found to be conserved among PICP homologues. The two PICPs are the first reported enzymatic markers for late Iysosomes.

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Cloning and Heterologous Expression of Acetyl Xylan Esterase from Aspergillus ficuum

  • Jeong, Hye-Jong;Park, Seung-Mun;Yang, Mun-Sik;Kim, Dae-Hyeok
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.153-156
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    • 2000
  • Xylan, the major hemicellulose component of many plants, occurs naturally in a partially acetylated form and lignin, the most resistant component in plant cell wall degradation, is also attached to ${\beta}-1,4-linked-D-xylose$ backbone through the ester linkage. Esterases are required to release the esterified substituent and acetyl esterases are important in the complete degradation of acetylated polysaccharides, like pectins and xylans. The gene(Axe) encoding acetyl xylan estarase(AXE) was isolated from genomic ${\lambda}$ library from Aspergillus ficuum. Nucleotide sequencing of the Axe gene indicated that the gene was separated with two intervening sequences and the amino acid sequence comparison revealed that it was closely related to that from A. awamori with the 92 % indentity. Heterologous expression of AXE was conducted by using YEp352 and Saccharomyces cerevisae 2805 as a vector and host expression system, respectively. The Axe gene was placed between GAL1 promoter and GAL7 terminator and then this recombinant vector was used to transform S. cerevisiae 2805 strain. Culture filtrate of the transformed yeast was assayed for the presence of AXE activity by spectrophotometry and, comparing with the host strain, four to five times of enzyme activity was detected in culture filtrate of transformed yeast.

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Temperature-Pressure Estimation of Metasediments in Seosan Area (서산지역의 변성퇴적암류에 대한 온도-압력 추정)

  • Song, Yungoo;Moon, Hi-Soo;Lee, Han Yeang
    • Economic and Environmental Geology
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    • v.22 no.4
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    • pp.371-379
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    • 1989
  • Peak or near-peak metamorphic temperatures and pressures for Seosan area could be estimated from major element method. Temperatures calculated from garnet-biotite geothermoneter(Ferry and Spear, 1978; Ganguly and Saxena, 1984) are $620{\pm}40^{\circ}C$ and $520{\pm}20^{\circ}C$ for Seosan and Daesan Formation respectively. Presures derived from garnet-plagiciase-$Al_2Si_O_2$-quartz geobarometer(Newton and Haselton, 1981; Ganguly and Saxena, 1984)suggest 5-6kb for both of Seosan and Daesan Formation. These results suggest that under isobaric, Seosan Formation underwent relatively high temperature metamorphism compared with Daesan Formation. Chemical zonations of garnet for major elements such as Fe, Mg, and Mn in Seosan and Dasan Formation show different patterns each other probably caused by different thermal history.

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Isolation of Oil-Degrading Bacterium, Providencia rettagei 4A3 and Characterization of Crude Oil Degradation (유류분해균 Providencia rettgeri 4A3의 분리 및 원유분해 특성)

  • 김상우;유주순;이상철;조영수;이영춘
    • Journal of Life Science
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    • v.11 no.6
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    • pp.530-536
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    • 2001
  • Several bacterial strains utilizing crude oil as their sole carbon and energy source were isolated from marine environment polluted by crude oil. Among them, the selected strain 4A3 showed strong degradation activity for crude oil. This strain was identified as a Provindencia rettgeri 4A3 based on the morphological, biochemical, and physiological characteristics. The optimum cultural conditions was as follows; 26$^{\circ}C$ for temperature and 7.0 for initial pH. Additionally, the optimal concentration of sodium chloride was 2.0%, indicating that this strain was derived from sea water. The emulsifying activity of 4A3 was the highest after 3 days of cultivation under the condition of 2.0% sodium chloride, pH 7.0 and 26$^{\circ}C$. This strain had one cryptic plasmid in 7.0kb size.

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