• The Journal of Korean Society of Virology
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• v.28 no.4
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• pp.383-391
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• 1998

A murine monoclonal antibody (mAb) specific for the envelope glycoprotein gp120 of human immunodeficiency virus type-I (HIV -1) was chemically coupled to pokeweed antiviral protein (PAP) from Phytolacca americana. The immunotoxin was purified by FPLC using S200 colum. The purified immunotoxin efficiently bound to HIV-infected T cells as evidenced by fluorescenceactivated cell sorter analysis. The immunotoxin selectively killed human T lymphoid lines infected with $HIV-1_{IIIB}$ at less than 250 pM of the immunotoxin cells, while PAP or mAb alone did not have any significant effect on infected cells. The uninfected control T cell lines were not affected. Human cells infected with HIV-2 or other HIV-1 strains were not killed, suggesting that the killing depends completely on the antibody used for coupling. These in vitro results suggest that the PAP-mAb conjugate may be used to selectively remove cells expressing viral antigens from individuals infected with HIV.

• Journal of the Korean Society of Tobacco Science
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• v.23 no.2
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• pp.123-132
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• 2001

In order to transform pokeweed antiviral protein cDNA to tobacco plant, total RNA was extracted from Phytolacca americana. PAP-II cDNA was synthesized from purified total RNA via RT-PCR and subcloned to recombinant vector pBluescript II SK-. 10 deletion mutant PAP-II cDNA fragments which were sequentially deleted from N-terminal by 90bp were synthesized from PAP-II cDNA except leading frame by PCR with primers designed in our laboratory. To select non-cytotoxic clone, pAc55M was constructed with yeast expression vector pAc55 and multicloning site(MCS). Sequentially deleted mutant PAP-II cDNAs were cloned on downstream of gall promoter of pAc55M. 6 non-cytotoxic deletion mutant PAP-II cDNA were selected. Selected cDNAs were cloned into plant expression vector pKGT101BH for transformation of these clones to plant through Agrobacterium tumefacience. After cloning, recombinant pKGT101BH carrying deleted mutant PAP-IIcDNA were transformed to Nicotiana tabacum cv. NC567. Transformed tobacco plants cultured on shooting and rooting media were transfered to green-house. About four weeks later, these plants were infected with physically infection using carborandum with PVY-VN strain. After 4 weeks, plants resistant to virus were selected , and seeds of these plants were gathered. Southern blot hybridization showed deleted fragments by 220bp and 420bp, so resistant ability of these plants is due to mutant PAP-II cDNA.

• Journal of the Korean Society of Tobacco Science
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• v.21 no.1
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• pp.57-63
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• 1999

Pokeweed antiviral protein II (PAP-II) encoding cDNA was synthesized by reverse-transcriptase polymerase chain reaction (RT-PCR) from Phytolacca american a leaf. The PAP-II cDNA fragment of 974bp was subcloned to pBluescript II SK- SmaI site and the inserted PAP-II cDNA fragment was sequenced by dideoxy sequencing method. The number of nucleotides of PAP-II cDNA coding region containing start and stop codon was 933bp. To develop a virus-resistant tobacco plant, PAP-II cDNA fragment was inserted to pKGT101B and the insertion of PAP-II cDNA fragment was confirmed by restriction enzyme analysis and colony PCR.

• Journal of Life Science
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• v.6 no.2
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• pp.87-93
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• 1996

귀화식물인 미국자리공의 군락 구조와 무기환경요인을 추적하였다. 서식지 식생은 Pinus densiflora, persicaria perfoliata, Miscanthus sinensis, Smilax china, Rubus crataegifolius, Paederia scandens, Rosa multiflora, Commelina communis, Carex lancelolatl가 우점 하였으며 파괴된Pinus densiflora 아극상 감림군략에 침임하여 적응한 형태였다. 서식지 토양의 속성은 pH 4.6의 높은 산도, 높은 중금속과 유기물 함량이 특징적이었으며 이는 전형적인 황지식물군락의 환경요인이었다. 생장조절실 내에서 수행한 제한요인 처리에 의해 나타나는 유식물의 피해반응은 인공산성비에 의한 pH2.0과 500Lux의 광도에서 나타났으며 일시위조점은 pF2.6-2.7에서, 그리고 영구위조점은 pF2.8-2.9에서 각각 나타났다.

• Archives of Pharmacal Research
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• v.18 no.4
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• pp.267-270
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• 1995

We examined the chronic effect of dextromethorphan(DM) on the cellular immune responses in mice. T cell simulator, phytohemagglutinin did not show singificant effect on lymphocyte proliferation. Costimulator of T and B cell, pokeweed mitogen, and B cell stimulator, lipopolysaccharide exhibited DM-induced decreased lymphocyte proliferation. Singificantly suppressed natural killer (NR) cell cytotoxicity was evidenced following 6 months DM exposure. These results suggest that chronic DM administration pertub B cell functioning and NK cell cytotoxicity. In addition, prenatal DM exposure did not potentiate the immunomodulation in postnatal effect induced by chronic DM.

• Journal of Preventive Medicine and Public Health
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• v.27 no.1 s.45
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• pp.11-24
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• 1994

The studies reported here were undertaken to investigate the effects of mercury chloride on immune system of Balb/c mouse employing a flexible tier of in vitro and in vivo assays. Mercury chloride inhibited the proliferative responses of spleen cells to lipopolysaccharide, pokeweed mitogen, and phytohemagglutinin as a dose-dependent manner. This inhibitory effect was observed not only when $HgCl_2$ was added 2nd or 3rd day of 3 days culture period but also when spleen cells was pretreated with $HgCl_2$ for 2 hours. Mercury chloride, however, potentiated the production of IgM and IgG from spleen cells. During the $HgCl_2$ administration by drinking for 3 weeks, the weight gain of mice was significantly blunted than that o control group mice, while no overt signs related to mercury toxicity were noted in any mice of experimental group. There was no change in thymus and spleen weights, and in histological findings of kidney, bone marrow of femur, thymus, spleen, and popliteal lymph node after 3 weeks of mercury exposure. However, $HgCl_2$ induced a significant increase of total serum IgM, IgG including $IgG_1,\;IgG_{2a}\;and\;IgG_{2b}$, and IgE in Balb/c mice. Treatment in vivo with anti-IL-4 monoclonal antibody significantly abrogated the $HgCl_2$-induced increase in total serum IgG1 and IgE. Whereas $HgCl_2$ potentiated total serum IgM and IgG, there was no difference in total serum hemagglutinin to SRBC (Sheep Red Blood Cell) between experimental and control group mice when these mice were immunized with SRBC. All these findings observed in Balb/c mice suggest that mercury perturbates well-orchestrated regulation of immune responses before developing histopathological changes in lymphoid tissues.

• Korean Journal of Medicinal Crop Science
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• v.9 no.2
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• pp.156-165
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• 2001

Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.42 no.2
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• pp.153-159
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• 1997

Pokeweed, a polycarpic plant, has been used as herbage medicine, vegetable or dye. It, however, is known as an aggressive plant in the vicinity of the industrial area evolving air pollutants. The experiment was done to determine the effect of priming using nitrates, germination temperature and light quality on germination of its seed to get information on the optimum germination process as well as its establishment. The daily percent germination was measured to 12 days after sowing since its seeds were treated by two different nitrates [KNO$_3$, Ca(NO$_3$)$_2$]. their different concentrations (0, 50, 150, 300mM), then treatment duration (1, 3, 6 days), different germination temperature (day /night; 30/30, 30/20, 20/30, 20/2$0^{\circ}C$) and light quality (red, white, dark) before or during germination. The percent germination was greater in the KNO$_3$ treatment than in the Ca(NO$_3$)$_2$ but in the priming treatment with KNO$_3$ in comparison with no-priming. In the priming treatment with KNO$_3$, the percent germination was increased with its increased concentration to 150mM although decreased with delayed duration to 6 days. Regardless of light quality, the greater percent germination was shown in the order of 2$0^{\circ}C$ constant and 20/3$0^{\circ}C$ alternative, 3$0^{\circ}C$ constant, 30/2$0^{\circ}C$ alternative temperature. The germination was less in the dark during germination than in the illumination in which the red light had greater percent germination compared to white light. The seeds primed with KNO$_3$ were germinated under the alternative temperature even in the dark condition.

• The Korean Journal of Ecology
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• v.20 no.1
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• pp.43-49
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• 1997

Effects of addition of soil ameliorants on the growth and nutrient absorption of Zea mays, Miscanthus sinensis and Phytolacca americana were investigated. Acid soil from Yeocheon Industrial Complex were used for the control plot (AS). We made two treatment plots, acid soil + lime (AS + L) and acid soil + dolomite (AS+D). Ration of acid soil : soil ameliorants in freatment plot was 50 : 1(V:V). Acid soil and soil ameliorants were mixed thoroughly before use. Shoot length of corn in AS+L and AS+D was considerably increased by 141% and 137%, respectively, compared with that in AS. Pokeweed in AS+L and AS+D also increased by 183% and 152%, respectively, compared with that in AS. However, growth of Miscanthus sinensis showed slight difference between the control and the treatment plots. Biomass of corn and porkweed in the treatment plots were also greater than those in the control plots. During the growth experiment with corn, concentrations of Ca and Mg in soil were increased and A1 decreased with increased soil pH in the treatment plots. Amount of aluminum absorbed by corn in the control plot was greater than that in the treatment plots. In case of Miscanthus sinensis, however, aluminum absorption in the control plot was lower than those in the treatment plots.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.257-263
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• 2011

In general, the blood cell culture is a common method for animal chromosome preparation. However, every animal and its cells have unique physiological characteristics and functions. Hence, it is very difficult to find the suitable method of chromosome preparation using animal lymphocyte culture. This study was carried out to fine the suitable method of chromosome preparation using lymphocytes cultures in mammalians and aves including cattle, rat, mouse and chicken. To seek the optimal method of lymphocyte culture in each animal, $2^3$ factorial experiment was designed. The design evaluated three main effects in culture duration, kinds of mitogen supplements and colcemid exposure time with two levels within each effect. The mitotic index and the score of chromosome morphology were analyzed. In results, the suitable methods of lymphocyte culture for chromosome preparation were 72 hours culture, pokeweed mitogen(PWM) supplement and 90 minutes of colcemid exposure in cattle, 72 hours culture, PWM supplement and 50 minutes of colcemid exposure in chicken, 96 hours culture, concanavalin A supplement and 90 minutes of colcemid exposure in rat, and 72 hours culture, PWM supplement and 50 minutes of colcemid exposure in mouse, respectively. In conclusion, kinds of mitogen, culture duration and colcemid exposure time significantly affected the mitotic index and chromosome morphology, in animal lymphocyte culture. The interaction effects between/among treatment factors were also statistically significant.