• Title/Summary/Keyword: blot

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Induction of Porcine Oocytes Maturation by MAP Kinase (MAP Kinase에 의한 돼지 난성숙의 유기)

  • 장규태;박미령;윤창현
    • Korean Journal of Animal Reproduction
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    • v.22 no.3
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    • pp.277-286
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    • 1998
  • The effect of MAP kinase activity on maturation of porcine oocytes was investigated. MAP kinase was detected by immunofluorescence staining in nucleus of oocytes just before entering GVBD (germinal vesicle breakdown)stage. In a Western blot with GV (germinal vesicle) from these oocytes (cultured for 25 hours), a shift of MAP kinase band a, pp.ared, suggesting an activated stage of the kinase. No activity was shown in the blot with GV isolated from, oocytes cultured for 0 hour. To confirm that activation of MAP kinase induce GVBD, we microinjected MAP kinase purified from matured oocytes starfish into the cytoplasm of oocytes in GV stage (cultured for 0 hour). The injected MAP kinase did not cause early a, pp.arance of GVBD. No oocytes showed GVBD state until 20 hours of culture. Activity of MAP kinase did not increase significantly after the injection. When the exogenous MAP kinase was injected into GV, GVBD was induced in about 20% of oocytes cultured for 5 to 10 hours. These results suggest that MAP kinase is traslocated to nucleus and function as a factor inducing GVBD.

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Isolation of Differentially Expressed Genes by Low Temperature Treatment in Winter Oilseed Rape (Brassica napus L. cv. Tammi)

  • Chun Jong-Un;Seo Dong-Joong;Bae Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.19 no.3
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    • pp.440-446
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    • 2006
  • To investigate flowering related genes in winter-type oilseed rape (Brassica napus L. cv. Tammi), differentially expressed genes were isolated from leaves of the plant after low temperature treatment which is requirements for floral induction. As a result of suppression subtractive hybridization (SSH), 288 clones were randomly selected from SSH library. Using reverse Northern blot analysis, 150 of 288 clones were identified to be differentially expressed. Out of these 150 clones, 45 clones showed very high identities with the known genes. Four clones showed very high identities over 90% with metallothionein-like gene that is related to flowering-induced genes. Of these 4 clones, the cDNA clone, rfs-13, revealed high identity with meotallothionein-like protein in Arabidopsis thaliana (98%) and Brassica compestris (89%). Furthermore, gene expressed in immature flower stages was confirmed by Northern blot analysis.

The effcect of Gorynju and a Soju extract Psoraleae fructus on Melanin synthesis of B16 melanoma cells (보골지(補骨脂)의 고량주(高梁酒), 소주(燒酒) 추출물이 B16 melanoma 세포주의 멜라닌 합성에 미치는 영향)

  • Seo, Hyeong-Sik
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.21 no.1
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    • pp.96-103
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    • 2008
  • Objective : This research was carried out to compare the effect of Gorynju and a Soju extract Psoraleae fructus on melanin synthesis of B16 melanoma cells. Methods : To investigate melanin synthesis of B16 melanoma cells, this research was measured cell survival, tyrosinase activity, melanin synthesis, western blot. Results : Both Gorynju and Soju extract Psoraleae fructus, cell toxicity depended on the density. Tyrosinase activity depended on the density of Gorynju extract Psoraleae fructus and statistic was showed significant(0.5, 1, 2, 3 ${\mu}g/ml$), in a Soju extract Psoraleae fructus, 1 ${\mu}g/ml$ were showed significant. Melanin synthesis was showed significant in a Soju extract Psoraleae fructus(3, 4 ${\mu}g/ml$). Western blot was showed to depend on the density of Gorynju and a Soju extract Psoraleae fructus. Conclusions : In a tyrosinase activity and a melanin synthesis, the intermediate alcohol of Gorynju and a Soju may be suitable to use.

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cis-Diamminedichloroplatinum (II) (CDDP) Inhibits Bluetongue Virus (BTV) Core Associated Transcriptase Activity (CDDP를 처리한 Bluetongue Virus Core의 전사저해)

  • ;Manning, JaRue S.
    • Korean Journal of Microbiology
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    • v.29 no.1
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    • pp.34-39
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    • 1991
  • The BTV core associated transcriptase activity, assayed by acid precipitable counts, was reduced to an undetectable level after treat the core with .$100{\mu}{\M}$ CDDP. When the RNA transcripts prepared from the CDDP treated BTV core were analysed on agaroseurea gel, it was observed that the band intensity of the large size RNA was reduced while the band intensity of the small size RNA was enhanced. Northern blot analysis showed that much of the small size RNAs appeared to be prematurely terminated transcripts. These results suggest that CDDP adduction to the template RNA blocks chain elongation process of the virion bound transcriptase that is ultimately responsible for the inactivation of BTV infectivity.

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Tissue-Specific Regulation of Angiotensinogen and Angiotensin II Receptor Gene Expression in Deoxycorticosterone Acetate-Salt Hypertensive Rats

  • Lee, Jong-Un;An, Mi-Ra
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.3
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    • pp.315-320
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    • 1999
  • Molecular regulation of the renin-angiotensin system (RAS) was investigated in deoxycorticosterone acetate (DOCA)-salt hypertension. The expression of renin, angiotensinogen and angiotensin II receptor genes in the kidney and liver was determined by Northern blot analysis in rats which were made DOCA-salt hypertensive over the period of 2 or 4 weeks. Along with the hypertension, renin mRNA was decreased in the remnant kidney. The expression of angiotensinogen gene was not significantly altered in the kidney, but was significantly decreased in the liver. The expression of angiotensin II receptor gene was increased in the kidney, while it remained unaltered in the liver. The duration of hypertension did not affect the altered gene expression. It is suggested that the components of RAS are transcriptionally regulated in DOCA-salt hypertension in a tissue-specific manner.

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Identification and Characterization of Nitric Oxide Synthase in Salmonella typhimurium

  • Choi, Don-Woong;Oh, Hye-Young;Hong, Sung-Youl;Han, Jeung-Whan;Lee, Hyang-Woo
    • Archives of Pharmacal Research
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    • v.23 no.4
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    • pp.407-412
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    • 2000
  • The presence of the nitric oxide synthase (NOS) enzyme from Salmonella typhimurium (S. typhimurium) was identified by measuring radiolabeled L-$[^3H]$citrulline and NO, and Western blot analysis. NOS was partially purified by both Mono Q ion exchange and Superose 12HR size exclusion column chromatography, sequentially. The molecular weight of NOS was estimated to be 93.3 kDa by Western blot analysis. The enzyme showed a significant dependency on the typical NOS cofactors; an apparent Km for L-arginine of 34.7 mM and maximum activity between $37^{\circ}C$ and $43^{\circ}C$. The activity was inhibited by NOS inhibitors such as aminoguanidine and $N^{G}$ $N^{G}$-dimethyl-L-arginine. taken together, partially purified NOS in S. typhimurium is assumed to be a different isoform of mammalian NOSs.OSs.

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Inheritance and Expression of Antisense Polygalacturonase Gene in Transgenic Tomato (Antisense Polygalacturonase 유전자 형질전환 토마토의 후대 발현 분석)

  • 김영미;한장호;김용환;이성곤;황영수
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.2
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    • pp.131-134
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    • 1998
  • $\textrm{T}_{5}$ progeny of one transgenic tomato line (To9) carrying antisense polygalacturonase (PG) cDNA was generated by selfing. Five $\textrm{T}_{5}$ plants were used to analyse in detail. The PG antisense gene was stably inherited through fifth generations. In all five $\textrm{T}_{5}$ plants, expression of the antisense transcripts were detected. In consequence, it led to a reduction of the PG enzyme activity in ripe fruit to between 37% and 65% that of normal. In two plants the expression of endogenous PG gene was inhibited in ripe fruit.

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Expression of Proteinase Inhibitor II gene in Transgenic Flowering Cabbage, Brassica oleracea var. acephala DC. (형질전환된 꽃양배추에서 Proteinase Inhibitor II 유전자의 발현)

  • 김창길;정재동
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.2
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    • pp.95-98
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    • 1998
  • Hypocotyl explants of flowering cabbage were cocultured with Agrobacterium tumefaciens LBA4404;;pGA875 harboring proteinase inhibitor II(PI-II) cDNA and then regenerated into plants. Sucessful transcripts of PI-II gene were detected by RNA dot blot analysis. Bioassay was conducted on transgenic flowering cabbage. It was confirmed that insecticidal activities of transformants were much higer than that of control plants. In progeny test of hansformants, 27.4% of T$_1$ seeds was resistant on MS medium containing 20 mg/L kanamycin.

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The Expression Characterization of Chicken Uncoupling Protein Gene

  • Zhao, Jian-Guo;Li, Hui;Wang, Yu-Xiang;Meng, He
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.11
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    • pp.1552-1556
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    • 2005
  • The UCPs are members of the mitochondrial inner membrane transporter family, present in the mitochondrial inner membrane. Their main function is increasing the energy expenditure via diminishing the resulting production of ATP from mitochondrial oxidative phosphorylation instead of yielding dissipative heat. They are associated with the metabolism of fat and regulation of energy expenditure. The UCP gene can be viewed as the candidate gene for chicken fatness. In the present study, RT-PCR and Northern Blot methods were developed to investigate the expression of the UCP gene in ten tissues including heart, liver, spleen, lung, kidney, gizzard, intestine, brain, breast muscle and abdominal fat of chicken. The results of both RT-PCR and Northern Blot methods showed that the UCP gene expressed specific in breast muscle. The expression levels of UCP gene in breast muscles from egg-type and meat-type chickens of hatching, 2, 4, 6 and 8 wk of age were detected by RT-PCR assay and results showed that the expression levels of UCP gene were related to breeds. Expression level of UCP gene in layers was higher than that in broilers at various weeks of age except at 6 wk. The UCP gene's expression was higher at 6 wk and had no significant difference among other weeks of age in broilers; in layers the expression level of UCP gene had no significant difference among weeks of age. The experiment results also showed that insulin could increase the expression level of UCP gene by 40% compared with control group.