• 제목/요약/키워드: DNA incorporation

검색결과 166건 처리시간 0.021초

유선발달에 있어서 cAMP, EGF, IGF-I 및 단백질 인산화 작용의 역할 I. EGF, IGF-I 및 Photoreactive Cyclic AMP가 유선상피세포의 DNA합성에 미치는 효과 (Role of cAMP, EGF, IGF-I and Protein Phosphorylation in Mammary Development I. Effect of EGF, IGF-I and Photoreactive Cyclic AMP on DNA Synthesis of Mammary Epithelial Cell)

  • 여인서;박춘근;홍병주
    • 한국가축번식학회지
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    • 제17권1호
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    • pp.49-56
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    • 1993
  • Mouse mammary epithelial cells(NMuMG) were plated onto 24 well phates(100,000 cells/well), in DMEM supplemented with 10% fetal calf serum. After serum starvation for 24 hours, EGF)0~100ng/ml) was added simultaneously with IGF-I(10ng/ml), 1$\mu$M photoreactive cAMP(4,5-dimethoxy-2-nitrobenzyl adenosine-3',5' cyclic monophosphate, DMNB) or IGF-I plus DMNB. After 2 hours, the cells were expposed to UV light(300nm, 3 second pulse0 in order to activate DMNB which induces a rapid transient increase in intracellular cAMP upon UV irradiation. DNA synthesis was estimated as incorporation of 3H-thymidine into DNA(1 hour pulse with 1$\mu$Ci/ml, 18~19 hours after UV exposure). Without IGF-I or DMNB, EGF(10 or 100ng/ml) increased DNA synthesis from 8,362 dpm/well in control to 16,345 or 18,684 dpm/well with EGF(pooled SE=1,239 dpm/well, P<0.05). IGF-I or IGF-I plus DMNB alone increased DNA synthesis from 8,362 dpm/well in control to 17,307 or 20,427 dpm/well, respectively(P<0.05). Addition of IGF-I, DMNB or IGF-I plus DMNB into 0~100ng/ml EGF did not significantly change the shape of dose response curve of EGF alone. In other experiment, EGF or IGF-I plus DMNB into 10ng/ml EGF group exhibited interaction effect in DNAsynthesis [EGF(10ng/ml)=18,497; IGF-I+EGF=22,837; DMNB+EGF=20,658 ; IGF-I+DMNB+EGF=29,658, pooled SE=1,055, P<0.05]. These results indicate that simultaneous activation of EGF, IGF-I and intracellular cAMP interact in DNA synthesis of mouse mammary epithelial cells.

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Matrix Attachment Regions (MARs) as a Transformation Booster in Recalcitrant Plant Species

  • Han, Kyung-Hwan
    • 식물조직배양학회지
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    • 제24권4호
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    • pp.225-231
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    • 1997
  • For genetic engineering to be commercially viable, an efficient transformation system is needed to produce transgenic plane from diverse genotypes ("generalized protocol"). Development of such a system requires optimization of a number of components such as gene transfer agent, plant tissues competent for both regeneration and transformation, and control of transgene expression. Although several novel gene transfer methods have been developed for plane, a majority of stably transformed plane express the introduced genes at low levels. Moreover, silencing of selectable marker genes shortly after their incorporation into plant chromosomes may result in low recovery of transgenic tissues from selection. Matrix attachment regions (MARs) are DNA sequences that bind to the cell's proteinaceous nuclear matrix to form DNA loop domains. MARs have been shown to increase transgene expression in tobacco cells, and reduce position in mature transgenic plants. Flanking an antibiotic resistance transgene with MARs should therefore lead to improved rates of transformation in a diversity of species, and may permit recalcitrant species and genotypes to be successfully transformed. Literature review and recent data from my laboratory suggest that MARs can serve as a transformation booster in recalcitrant plant species.

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Evaluation of Telomerase Inhibitors Using DE81 Filter Spotting Method from Natural Products

  • Lee, Sung-Jin;Woongchon Mar
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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    • pp.183-183
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    • 1998
  • Telomerase synthesizes telomeric DNA repeats onto chromosome ends de novo. Telomerase activation and telomere shortening in human somatic cells have been implicated in cell tumorigenesis and immortalization. In order to find the potential inhibitors against telomerase activitiy which can be used as potential anticancer agents, we screened about 100 kinds of natural products after partition into n-hexane, ethyl acetate and aqueous layers from methanol extracts. The inhibitory effects of these materials against telomerase enzyme activity were tested in 293T cell culture using telomeric repeat amplification protocol(TRAP). The incorporation of $\^$32/P-dGTP into amplified DNA was measured by adsorption to Whatman DE81 paper instead of using TRAP assay for screening the extracts of natural products. Strong effective compounds were not found in this study but DE81 filter spotting method may be a useful model for the screening. Some of the compounds which showed somewhat inhibitory effects had cytotoxic effects also.

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Human Cytomegalovirus 감염에 대한 파파베린과 뉴클레오사이드 유사체의 항바이러스 효과 (Antiviral Activity of Papaverine and Nucleoside Analogs on the Human Cytomegalovirus Infection)

  • 이찬희
    • 미생물학회지
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    • 제29권1호
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    • pp.25-33
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    • 1991
  • Antiviral activities of papaverine and nucleoside analogs, 9-[(1,3-dihydroxy-2-propoxy)methyl]guanine (DHPG) and acyclovir, against human cytomegalovirus (HCMV) infection were compared in vitro. Papaverine and DHPG were effective in reducing infectious HCMV yields with $ED_{50}{\s}$ (effective dose 50: the concentraion at which 50% of virus yields was obtained) of approximately 1.02 and $0.45{\mu}{\M}$, respectively; while acyclovir was less effective with an $ED_{50}$ of about $10.4{\mu}{\M}$The relative cytotoxicity of these drugs was evaluated under the same conditions used to measure infectious HCMV yields. Papaverine and DHPG demonstrated little cellular toxicity as measured by their effect on the viability of confluent cells at concentrations in the range of those demonstrating potent inhibition of HCMV replication. Similarly, protein synthesis was largely unaffected by these drugs in stationary mock-infected cells as measured by the incorporation of isotopically labelled amino acids. In contrast, cellular DNA synthesis was invariably reduced in the presence of either drug. HCMV-specific DNA synthesis was also strongly inhibited by papaverine and DHPG.

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DHA가 풍부한 어유가 새끼쥐의 뇌발달과 학습능력에 미치는 영향 (Effect of DHA-Rich Fish Oil on Brain Development and Learing Ability in Rats)

  • 정경숙
    • Journal of Nutrition and Health
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    • 제29권3호
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    • pp.267-277
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    • 1996
  • Effect of DHA-rich fish oil on brain development and learning ability has been studied in Sprague Dawley rats. Female rats were fed experimental diets containing either corn oil fish oil at 10%(w/w) level throughout the gestation and lactation. Corn oil was added in fish oil diet to supply essential fatty acid at 2.3% of the calories. All male pups were weaned to the same diets of dams at 21-days after birth. Plasma fatty acid composition was analyzed for dams and pups at 21-days, 28-days and 22-weeks after birth. The analysis of DNA and fatty acid profile in the brain were undertaken at birth, 3, 7, 14, 21, 28 days and 22 weeks after birth and learning ability was tested at 18-20 weeks of age. Regardless of dietary fats, arachidonic acid(AA) and docosahexaenoic acid(DHA) were the principal polyunsaturated fatty acids in the brain. Rats fed CO diet showed a continouus increase of AA content in the brain from 10.9%(at birth) to maximum 15.3% level (14-days old), while the rars fed FO diet showed 78-79% of CO group throughout the period. Rats fed FO diet showed higher incorparation of DHA from 15.2% at birth to a maximum level of 18.5% at 140days, while the rats fed CO diet showed only 7.0% incorporation of DHA at birth and a maximum level of 11.1% at 21-days. Compared to CO group, FO group showed lower ratio of chol/PL and higher content of DHA in brain microsomal membrane, resulting in better membrane fluidity. Total amount of DNA per gram of brain was reached maximum level at 21 days in both groups. This would be a period of the cell proliferation during brain development. Overall, the rats fed fish oil diet showed a higher incorporation of DHA and membrane fluidity in the brain and better learning performances (p<0.05).

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가잠의 휴면성에 관한 세포학적 연구 (Cytological Studies of Diapause in the Silkworm, Bombyx mori L.)

  • Park, Kwang E.
    • 한국잠사곤충학회지
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    • 제18권2호
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    • pp.1-60
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    • 1976
  • 본 연구에서는 전자현미경과 Radioautography에 의하여 가잠의 휴면 기구를 연구하였다. 1. 가잠의 휴면성은 최청중의 온도와 명장에 의하여 변하기 쉬운 것으로서 특히 배자의 후기와 관계가 깊다는 사실은 잘 알려저 있다. 그러므로 저온과 고온최청의 경우 가잠의 배자 발생 단계중에 있어서 핵산과 단백질의 전구물질을 사용하여 합성의 Pattern을 조사하였다. 고온최청 때에는 반전후 3일째에 $^3$H-glycine이 배자의 뇌와 식도하신경구에 다량으로 들어가 있는 동시에 타부위에도 약간이나마 들어가 있었다. 그러나 저온최청 때에는 뇌와 식도하신 경구 그리고 타부위 사이에 차이가 없었다. 고온과 저온과의 사이에는 배자의 DNA 합성의 차리를 인정할 수 없었다. 산난후 20일째의 휴면난에 있어서는 소수의 중배엽세포의 핵에 들어가 있었으나 월연한 잠난에 있어서는 중배엽의 전핵에 들어가 있었다. 고온과 저온최청때에 반전후의 배자의 전부 즉 뇌와 식도하신경구의 주위에만 $^3$H-thymidine이 들어가 있는 것을 볼 수 있었으나 후부에서는 전혀 볼 수 없었다. 한편 전 최청기간중 또는 산난후 20일째의 난황세포에 있어서는 단백질합성과 나NA 합성을 인정하지 못하였으나 월년난 배자의 부속지 발생전까지는 RNA합성이 관찰되었다.

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불화나트륨이 조골세포의 생리적 활성에 미치는 영향 (THE EFFECT OF SODIUM FLUORIDE ON THE PHYSIOLOGICAL ROLE OF OSTEOBLASTIC CELL)

  • 김대업
    • 대한소아치과학회지
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    • 제25권3호
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    • pp.635-648
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    • 1998
  • The clinical use of fluoride with a well known osteogenic action in osteoporotic patients is rational, because this condition is characterized by impaired bone formation. However, its anabolic effect has not been demonstrated well in vitro. The purpose of this study was to investigate the effects of sodium fluoride on the physiological role of osteoblastic cell. Osteoblastic cells were isolated from fetal rat calvaria. The results were as follows : 1. Mineralized nodules were shown in osteoblastic cell cultures, which had been maintained in the presence of ascorbic acid and ${\beta}-glycerophosphate$ up to 21 days. When cultures were treated with pulses of 48 hr duration before apparent mineralization was occurring, 2-fold increased in their number was detected. 2. Alkaline phosphatase activity of osteoblastic cells was inhibited by sodium fluoride in dose dependent manner. 3. The effect of sodium fluoride on the osteoblastic cell proliferation was measured by the incorporation of $[^3H]$-thymidine into DNA. As a result, sodium fluoride at $1{\sim}100{\mu}M$ increased the $[^3H]$-thymidine incorporation into DNA in a dose dependent manner. 4. The signaling mechanism activated by sodium fluoride dose-dependently enhanced the tyrosine phosphorylation of the adaptor molecule $Shc^{p66}$ and their association with Grb2, one of earlier events in a MAP kinase activation pathway cascade used by a significant subset of G protein-coupled receptors. 5. The phosphorylation of CREB(cAMP response element binding protein)was inhibited by the sodium fluoride in MC3T3E1 cells. In conclusion, the results of this study suggested that the mitogenic effect of the sodium fluoride in MC3T3E1 cell was stimulated in a dose-dependent manner and suggested "an important role for the interaction between She and Grb2" in controlling the proliferation of osteoblasts.

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고려인삼이 마우스의 악하선 DNA 합성능에 미치는 영향( I ) (Influence of Panax Ginseng on DNA Synthesis of Submandibular Gland in Mice)

  • 권영진;채유병;장원상
    • The Korean Journal of Physiology
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    • 제8권1호
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    • pp.23-26
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    • 1974
  • 인삼주정추출액 투여에 의하여 마우스 악하선 조직의 DNA 합성능이 어떤 영향을 받는지를 알기 위하여 30마리의 마우스 (몸 무게 $18{\sim}20$ gm) 수컷을 15마리씩 두 무리로 나누어 한 무리에는 생리적 식염수를 다른 무리에는 인삼주정추출액(인삼주정추출물 4mg 을 생리적 식염수 1ml 속에 용해시켜 만든 용액)을 마우스의 몸 무게 10gm에 대하여 0.05ml씩 5일 동안 주사한 후 $[^3H]-thymidine$을 복강 속에 주사하고 1, 10 및 24시간 만에 도살하여 자기방사법을 이용하여 악하선의 합성능을 비교 관찰한 바 인삼주정추출액을 투여받은 마우스의 악하선 조직의 표지 세포수는 식염수만을 투여받은 마우스의 그것에 비하여 적었다. 그러므로 인삼주정추출액은 마우스의 악하선 조직 DNA 합성능을 어느 정도 억압한다고 믿어진다.

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Synthesis and Characterization of Oligonucleotides Containing Site-Specific Bulky $N^2$-Aralkylated Guanines and $N^6$-Aralkylated Adenines

  • Moon, Ki-Young;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • 제23권2호
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    • pp.139-146
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    • 2000
  • 7- Bromomethylbenz[a]anthracene is a known mutagen and carcinogen. The two major DNA adducts produced by this carcinogen, i.e., $N^2$-(benz[a]anthracen-7-yl methyl)-2'-deoxyguanosine (2, b[a]$a^2$G) and $N^6$-(benz[a]anthracen-7-ylmethyl)-2'-deoxyadenosine (4, b[a]$a^6$/A), as wel 1 as the simpler benzylated analogs,$N^2$-benzyl-2'deoxyguanosine (1, $bn^2$G) and $N^6$-benzyl-2'-deoxyadenosine (3, $bn^6$/A), were prepared by direct aralkylation of 2'-deoxyguanosine and 2'-deoxyadenosine. To determine the site-specific mutagenicity of these bulky exocyclic amino-substituted adducts, the suitably protected nucleosides were incorporated into 16-base oligodeoxyribonucleotides in place of a normal guanine or adenine residues which respectively are part of the ATG initiation codon for the lac Z' \alpha-complementation gene by using an in situ activation approach and automated phosphite triester synthetic methods. The base composition and the incorporation of the bulky adducts into synthetic oligonucleotides were characterized after purification of the modified oligonucleotides by enzymatic digestion and HPLC analysis.

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