• Title/Summary/Keyword: in-vitro

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Studies on the Standardization of Chinese Cabbage Kimchi (배추김치의 표준화 연구)

  • Cho, Eun-Ju;Lee, Seon-Mi;Rhee, Sook-Hee;Park, Kun-Young
    • Korean Journal of Food Science and Technology
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    • v.30 no.2
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    • pp.324-332
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    • 1998
  • In order to standardize the chinese cabbage kimchi, the preparation method, kinds of ingredients and levels of the ingredients were determined by the statistical survey of literatures obtained from cooking books, scientific papers and kimchi manufacturing factory. The standardized ingredient kinds and ratio of chinese cabbage kimchi were $13.0{\pm}7.0$ of radish, $2.0{\pm}0.5$ of green onion, $3.5{\pm}0.8\;or\;2.5{\m}0.3$ of red pepper powder, $1.4{\pm}0.4$ of garlic, $0.6{\pm}0.3$ of ginger, $2.2{\pm}1.6$ of anchovy juice, and $1.0{\pm}0.3$ of sugar in the proportion of 100 salted chinese cabbage, and the final salt concentration was adjusted to 2.7% using salt. Red pepper powder level was quite different from the literature sources, so sensory evaluation, chemical properties and antimutagenic effect and growth inhibitory effect on human cancer cells of the kimchi samples were studied to decide the proper ratio of the red pepper powder as an ingredient. Red pepper powder 3.5% (average level for kimchi manufacturing factory) added kimchi was better in quality than red pepper powder 2.5% (average level for cooking books and scientific papers) added kimchi in sensory evaluation and chemical properties. The juice of red pepper powder 3.5% added kimchi showed not only the stronger antimutagenicity against aflatoxin $B_1$ in Salmonella typhimurium TA100 but also the higher inhibitory effect on the growth of AGS human gastric adenocarcinoma cells in SRB assay than that of red pepper powder 2.5% added kimchi. In conclusion, the standardized ratio of the ingredients was 13.0 radish, 2.0 green onion, 3.5 red pepper powder, 1.4 garlic, 0.6 ginger, 2.2 anchovy juice, 1.0 sugar, and 2.7 final salt concentration in the proportion of 100 salted chinese cabbage.

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The Ability of Cervus Elaphus Sibiricus Herbal Acupuncture to Inhibit the Generation of Inflammatory Enzymes on Collagen-induced Arthritis Mice (녹용약침(鹿茸藥鍼)이 CIA 모델 생쥐의 염증인자 생성억제에 미치는 영향)

  • Hwang, Jong-Soon;Hwang, Ji-Hye;Lee, Hyun-Jin;Lee, Dong-Gun;Kang, Min-Joo;Back, Song-Ook;Cho, Hyun-Seok;Kim, Kyung-Ho;Kim, Kap-Sung
    • Journal of Acupuncture Research
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    • v.24 no.6
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    • pp.1-14
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    • 2007
  • Backgrounds : Rheumatoid Arthritis(RA) is known as the chronic inflammatory diseasethat induces persistent inflammation in the joint cavity. The destruction of cartilage occurs as the result of bones destoyed by pannus, several influential cytokines induced by the synovial capsulitis, varieties of proteinases, $O_2$ radicals, and the secondary degenerative changes of articular cartilage. The type 2 collagen-induced arthritis model is used in recent experimental research on rheumatoid arthritis. Cervus elaphus sibiricus (Nockyong) has the effect of relieving pain by nourishing the muscles, joints, and bones. It is also known to be efficacious in promoting and enhancing the immune system. The objective of this study was to investigate the effect of Cervus elaphus sibiricus herbal acupuncture to inhibit the generation of proinflammatory enzyme on type 2 collagen-induced arthritis. I investigated the inhibition of mRNA transcription of MIF(macrophage migration inhibitory factor), $TNF-{\alpha}$(Tumor necrosis $factor-{\alpha}$) and MMP-9 (matrix metalloproteinase-9) of Cervus elaphus sibiricus herbal acupuncture using an in vitro test. Also investigated was the inhibition of differentiation of Th 1 cells and activation of cytokines(MIF, $TNF-{\alpha}$, IL-6, MMP-9), which are known to cause initial RA ,and are also related to the morphology of the synovial membranes of the joint capsule, by an in vivo test, using CIA(collagen induced arthritis) model mice. Materials & methods : The laboratory animals used in this experiment were 4 week-old DBA female mice, weighing approximately 20 grams, and adjusted to the laboratory environment. The experiment was divided into the normal group(NOR)-no treated group, control group(CON)-CIA induced group, and sample group(SAM)-Cervus elaphus sibiricus herbal acupuncture treated group. RA was induced in the mice via injection of $50{\mu}{\ell}$ C II mixed CFA. The Cervus elaphus sibiricus herbal acupuncture solution was applied on $GB_{35}$(陽陵泉) for 26 days from the 3rd day of RA inducement. The concentration of the solution was determined via a MTT assay. To research the effect on the expression of MIF, $TNF-{\alpha}$ and MMP-9 mRNA, RT-PCR was performed on synovial membrane cells from the knee joint of CIA mice. C II induced RA knee joint's histo-chemical synovial membrane was observed using a specimen model via the Hematoxilin and Eosin dying technique. Results : The expression of mRNA of RA-related cytokines such as MIF, $TNF-{\alpha}$, and MMP-9 dosedependently decreased in the cell from the synovial membranes of the joint, which is treated with Cervus elaphus sibiricus herbal acupuncture solution. In mice treated with Cervus elaphus sibiricusherbal acupuncture, the damage of synovial membranes of the joint was lessened, and differentiation of Th 1 cells was suppressed. The activation of RA-related cytokines such as MIF was suppressed, and the generation of $TNF-{\alpha}$ and MMP-9 showed a statistically significant decreas. Conclusions : It is speculated that Cervus elaphus sibiricus herbal acupuncture has the therapeutic effect of palliating the damage of the tissue impaired by RA by inhibition of the initial RA progression and by regulating excessive differentiation of Th 1 cell as it suppresses the generation of RA-related cytokines during the highest stage of RA by acting on pro-inflammatory enzymes.

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Protective Effects of Perilla frutescens Britt var. japonica Extracts from Oxidative Stress in Human HaCaT Keratinocytes (HaCaT 피부각질세포에서 들깻잎 추출물의 산화적 스트레스에 대한 항산화 효과)

  • Ji, Na;Song, Jia-Le;Kil, Jeung-Ha;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.2
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    • pp.161-167
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    • 2013
  • The aim of this study was to investigate the protective effects of methanolic extract from perilla (Perilla frutescens Britt var. japonica) leaves (PLME) on oxidative injury from hydrogen peroxide ($H_2O_2$) in human HaCaT keratinoctyes. Cells were co-incubated with various concentrations (0~200 ${\mu}g/mL$) of PLME for 24 hr, and then exposed to $H_2O_2$ (500 ${\mu}M$) for 4 hr. $H_2O_2$ significantly decreased cell viability (p<0.05). However, PLME provided protection from $H_2O_2$-induced HaCaT cell oxidation in a dose-dependent manner. To further investigate the protective effects of PLME on $H_2O_2$-induced oxidative stress in HaCaT cells, the cellular levels of lipid peroxidation, and antioxidant enzymes (including superoxide dismutase (SOD), glutathione peroxidase (GSH-px) and catalase (CAT)) were measured. PLME decreased cellular levels of lipid peroxidation, and also increased the activities of antioxidant enzymes. In addition, the antioxidant activities of PLME were also determined by DPPH and hydroxyl (${\cdot}OH$) radical scavenging assay, and major antioxidant compounds of PLME were measured by colorimetric methods. DPPH and ${\cdot}OH$ radical scavenging activities of PLME increased in a dose dependent manner and was similar to the DPPH scavenging activity of ascorbic acid at 50 ${\mu}g/mL$; however PLME activities were stronger than ascorbic acid (50 ${\mu}g/mL$) in the ${\cdot}OH$ scavenging assay. The amounts of antioxidant compounds, including total polyphenolics, total flavonoids, and total ascorbic acid from PLME were $52.2{\pm}1.1$ mg gallic acid (GAE)/g, $33.7{\pm}4.7$ mg rutin (RUE)/g, and $17.0{\pm}0.5$ mg ascorbic acid (AA)/g, respectively. These results suggest that PLME has a strong free radical-scavenging activity and a protective effect against $H_2O_2$-induced oxidative stress in the keratinocytes.

Effects of Herbal Complex on Blood Glucose in Streptozotocin-induced Diabetic Rats and in Mice Model of Metabolic Syndrome (생약복합제의 Streptozotocin 유발 당뇨 및 대사성증후군 모델 동물에서의 혈당에 미치는 효과)

  • Park, Han-Seok;Lee, Yeon-Sil;Choi, Se-Jin;Kim, Jin-Kyu;Lee, Yun-Lyul;Kim, Hyun-Gwen;Koo, Sam-Hoi;Ku, Dae-Hoy;Ki, Seung-Il;Lim, Soon-Sung
    • Korean Journal of Pharmacognosy
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    • v.40 no.3
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    • pp.196-204
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    • 2009
  • This study was carried out to investigate the in vivo and in vitro inhibitory effect of a traditional herbal complex (HC) extract prepared from a mixture of four oriental herbs (Dioscorea Rhizoma, Glycine soja Sieb. et Zucc, Bombycis corpus, Fermented Glycine soja) that have been widely used for the treatment and prevention of diabetes mellitus on hyperglycemia. The water extract of HC showed potent inhibitory effect on $\alpha$-glucosidase with $IC_{50}$ value of 1.24 mg/mL. Additionally, the ethanol extract of HC was also found to exhibit significant inhibitory effect against protein tyrosine phosphatase $1{\beta}$ ($PTP1{\beta}$), which is known as a major regulator of both insulin and leptin signaling. In the $PTP1{\beta}$ inhibitory assay, the most active n-hexane fraction obtained from the ethanol extract of HC, was identified as a mixture of fatty acid derivatives by gas chromatography-mass spectrometry (GC-MS). In high-fat diet-low dose streptozotocin (STZ)-induced diabetic rat, the water extract of HC improved the oral glucose intolerance as compared with rosiglitazone. HC also caused a marked decrease of body weight and fasting blood glucose and a significant improvement on glucose tolerance in metabolic syndrome mice model. These findings support that this traditional HC may be useful in the control of blood glucose in diabetes mellitus and metabolic syndrome.

Hepatitis B Virus-Induced TNF-a Expression in Hepa-lc1c7 Mouse Hepatoma Cell Line (마우스 Hepa-1c1c7 세포주에서 B형 간염 바이러스에 의한 tumor necrosis factor-a의 발현 유도)

  • Yea Sung Su;Jang Won Hee;Yang Young-Il;Lee Youn Jae;Kim Mi Seong;Seog Dae-Hyun;Park Yeong-Hong;Paik Kye-Hyung
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.38-44
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    • 2005
  • Infection with hepatitis B virus (HBV) is a major health problem worldwide. Although a tremendous amount has been known about HBV, there have been obstacles in the study of HBV due to the narrow host range of HBV limited to humans and primates. In the present study, we investigated the susceptibility to HBV infection of mouse hepatoma cell line, Hepa-1c1c7. In addition, based on that human hepatocytes infected by HBV increase the expression of the pro-inflammatory cytokine TNF-a, the inducibility of TNF-a expression by HBV in the cells was determined. HBV surface antigen (HBsAg) secretion was measured by the microparticle enzyme immunoassay and steady state mRNA expression was analyzed by quantitative competitive RT-PCR. Transient transfection of Hepa-1c1c7 cells with HBV expression vector resulted in a dose-dependent induction of TNF-a expression. Infection of Hepa-1c1c7 cells with the serum of HBV carrier also increased TNF-a mRNA expression. Both in the transfected and infected cells, HBV mRNA was expressed and significant HBsAg secretion was detected. There was no significant variation in $\beta-actin$ mRNA expression by HBV. These results demonstrate that HBV is infectious to Hepa-lc1c7 in vitro and the viral infection induces TNF-a expression, which suggests that Hepa-lc1c7, a mouse hepatoma cell line, may be a possible model system for analysis of various molecular aspects of HBV infection.

A Study on Organ Formation from the Tissues of Garlic and Onion In Vitro Culture (마늘의 유엽(幼葉) 및 양파의 인편배양(鱗片培養)으로부터 기관분화(器官分化)에 관한 연구(硏究))

  • Lee, Eun Mo;Lee, Yeong Bok
    • Korean Journal of Agricultural Science
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    • v.9 no.2
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    • pp.467-483
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    • 1982
  • Leaflets of Allium sativum L. (garlic) and leaf scales Allium cepa L. (onion) were cultured on the Murashige and Skoog(MS) medium to study the effects of plant growth regulators, temperature and light on the formation of shoot and root. And also, the potentiality of shoot or root formation of tissues in accordance with the leaf age and the region was discussed. In the experiment with garlic, the formation of shoot and root was more effective on the medium containing $BA\;2mg/{\ell}$ and $NAA\;1mg/{\ell}$ and it occured on the basal region of outer leaflets. The shoot formation was more effective at $20^{\circ}C$ than $28^{\circ}C$ under 16-hour daylength, and it was more effective in darkness than light condition at $28^{\circ}C$. The results of shoot and root formation on the new cloves of garlic harvested in 1982 were similar to those the old cloves of garlic harvested in 1981. In the experiment with leaf seales of onion, when the tissues of dormant leaf scales of onions harvested in 1981 were explanted on the MS medium, a few shoots were formed on the medium containing $BA\;2mg/{\ell}$ and $NAA\;1mg/{\ell}$. In this case,there were no differences on the shoot formation among the age and region of leaflets. On the culture of leaf scales, the shoot was not formed on the leaf scales located at outer or middle region, but it was formed on the medium containing BA accompanied with NAA at inner scales. On the medium complemented with BA alone, the tissues were not differentiated, but on the medium complemented with NAA, the callus was formed from tissues and then the root was formed through the callus.

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A STUDY ON MICROLEAKAGE OF PIT AND FISSURE SEALANT AFTER ENAMEL SURFACE TREATMENT (법랑질 표면처리방법에 따른 치면열구전색재의 미세누출에 관한 비교연구)

  • Hyun, Hong-Keun;Kim, Jung-Wook;Lee, Sang-Hoon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.3
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    • pp.512-521
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    • 2001
  • Recently. the reintroduction of air-abrasion technology in dentistry has added a new potential method of pretreating teeth prior to placing sealants. The purpose of this in vitro study was to investigate microleakages of several pit and fissure sealants following various surface pre-treatment methods to the enamel, because there was a question concerning the validity of claim that this method was better than the conventional acid etching. Permanent molars were divided randomly into nine groups and treated accordingly: acid etching(group 1, 4, 7), air-abrasion(group 2, 5, 8), acid etching after air-abrasion(group 3, 6, 9). Then the authors placed and cured the three kinds of sealants(group $1\sim3$: Teethmate-F, group $4\sim6$: Ultraseal XT plus, group $7\sim9$: Denseal) according to the manufacturer's order. They were observed to determine the degree of microleakage. And these specimens were separated from the corresponding enamels and examined by Scanning Electron Microscope. The following results were obtained: 1. In comparing microleakage among tooth surface treatment methods, air-abrasion group(group 2, 5, 8) showed the greatest microleakage, while combination(air-abrasion + acid etching) group showed the least (p<0.05). However, no significant differences were found between group 7 and 9. 2. The mean microleakages were ranked as follows(p<0.05): In acid etching groups, group 7> group 4> group 1. In air-abrasive groups, group 8>groups 5> group 2. In combination groups, group 9>group 3> group 6. However, no significant differences were found between group 4 and 7 between group 2 and 5 and between group 3 and 6. 3. SEM showed that comparably longer resin tags were distributed regularly in acid etching groups and that shorter ones irregularly in air-abrasion groups. It also showed that these two kinds of tags were distributed simultaneously in combination groups.

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Changes in Chlorophyll Contents of Leaves and pH of the Extracted Solutions from the Leaves of 7 Tree Species by pH Level (7개(個) 수종(樹種)의 잎이 pH수준별(水準別) 처리(處理)에 따른 엽록소(葉綠素) 함량(含量) 및 침출액(浸出液) pH변화(變化))

  • Woo, Jong Ho;Ahn, In Suk;Park, Young Goo
    • Journal of Korean Society of Forest Science
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    • v.87 no.2
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    • pp.145-152
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    • 1998
  • We conducted this study as a fundamental study on the response of various tree species against acid rain. The tree species used for this study were Zelkova serrata, Robinia pseudoacacia, Quercus acutissima, Prunus serrulata, Ginkgo biloba, Pinus koraiensis and Pinus densiflora. The leaves were examined for the pH changes by treatment time and the chlorophyll content into various pH solution in vitro. The results obtained were as follows ; 1. When the leaves were immersed in the solution of various pH(pH 3.0-pH 6.0) levels, the pH were changed to species specific pH ranges as pH 5.0~pH 5.5 of Z. serrata, pH 5.5~pH 6.0 of R. pseudoacacia, pH 4.5~pH 5.0 of Q. acutissima, pH 5.5~pH 6.0 of P. serrulata, pH 3.5~pH 4.5 of G. biloba, pH 3.5~pH 4.5 of P. koraiensis until 48 hours. However, in case of P. densiflora, it was difficult to find specific pH range of the species. Z. serrata, R. pseudoacacia and P. serrulata showed a little pH increasing by pH 2.0 solution treatment, while other species showed no change by the solution. 2. The amount of chlorophyll contents in Z. serrata, R. pseudoacacia and P. serrulata were decreased after immersing in the pH 2.0 solution. Chlorophyll content was almost constant in other pH levels. Other species showed almost constant chlorophyll contents in various pH levels and treatment time.

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Frequency of Micronuclei in Lymphocytes Following Gamma and Fast-neutron Irradiations (방사선 조사량에 따른 인체 정상 림파구의 미세핵 발생빈도)

  • Kim Sung-Ho;Cho Chul-Koo;Kim Tae-Hwan;Chung In-Yong;Yoo Seong-Yul;Koh Kyoung-Hwan;Yun Hyong-Geun
    • Radiation Oncology Journal
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    • v.11 no.1
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    • pp.35-42
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    • 1993
  • The dose response of the number of micronuclei in cytokinesis-blocked (CB) lymphocytes after in vitro irradiation with $\gamma$-rays and neutrons in the 5 dose ranges was studied for a heterogeneous population of 4 donors. One thousand binucleated cells were systematically scored for micronuclei. Measurements performed after irradiation showed a dose-dependent increase in micronuclei (MN) frequency in each of the donors studied. The dose-response curves were analyzed by a linear-quadratic model, frequencies per 1000 CB cells were ($0.31{\pm}0.049$)D+($0.0022{\pm}0.0002)D^2+(13.19{\pm}1.854) (r^2=1.000,\;X^2=0.7074,\;p=0.95$) following $\gamma$ irradiation, and ($0.99{\pm}0.528$)\;D+(0.0093{\pm}0.0047)\;D^2+(13.31{\pm}7.309)\;(r^2=0.996,\;X^2=7.6834,\;p=0.11) following neutrons irradiation (D is irradiation dose in cGy). The relative biological effectiveness (RBE) of neutrons compared with $\gamma$-rays was estimated by best fitting linear-quadratic model. In the micronuclei frequency between 0.05 and 0.8 per cell, the RBE of neutrons was $2.37{\pm}0.17$. Since the MN assay is simple and rapid, it may be a good tool for evaluating the $\gamma$-ray and neutron response.

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Distinct Spatio-temporal Expression Patterns of Patatin Promoter-GUS Gene Fusion in Transgenic Potato Microtubers (형질전환 감자 소괴경의 발달단계에 따른 Patatin Promoter-GUS 유전자의 발현 분석)

  • Youm, Jung-Won;Kim, Mi-Sun;Lee, Byoung-Chan;Kang, Won-Jin;Jeon, Jae-Heung;Joung, Hyouk;Kim, Hyun-Soon
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.13-18
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    • 2003
  • This study was carried out to investigate the expression patterns of foreign gene that controlled by tuber-specific patatin promoter in transgenic potatoes. Potato leaf disc cultured in vitro were transformed by the Agrobacterium strain LBA4404 containing pBl121 or pATGUS from potato cv. Desiree. In order to select the transgenic lines, gene-specific primers deduced from the NPTII were synthesized and used for polymerase chain reaction. The down part of the putative transgenic potatoes was transplanted weekly onto sucrose-enriched medium to accelerate the microtuber formation. RNA gel blot analysis was performed on the total RNAs obtained from tuber that had been harvested at a week interval. Also, histochemical assay was observed in the explants transformed with either pBI121 or pATGUS. Results showed that the transgenic plant containing pATGUS expressed GUS transcripts mainly at the tuber, not in stem, with the highest expression level in 5 weeks-grown microtubers. In contrast to pATGUS plants, the transformed plants with pBI121 showed an equal expression pattern throughout the whole developing stages. Consistent with RNA gel blot analysis, histochemical GUS staining and enzyme activity exhibited pATGUS transcripts were at the highest level in 5 weeks cultures. From these results, we suggest that the best stage to analyze the foreign gene introduced by patatin promoter into potato plants is at 5 weeks cultures after tuber formation.