• Title/Summary/Keyword: protein tissues

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A Study on Antioxidative Effects of Sipyimiguanjungtang and Osuyubujayijungtang, Korean Traditional Prescriptions for Soum Constitutes, in Brain and Liver of Rat (소음인(少陰人) 십이미관중탕(十二味寬中湯), 오수유부자이중탕(吳茱萸附子理中湯)이 흰쥐의 뇌(腦)와 간조직(肝組織)의 항산화(抗酸化) 기전(機轉)에 미치는 영향(影響))

  • Jung, Bong-yeon;Song, Il-byung
    • Journal of Sasang Constitutional Medicine
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    • v.11 no.2
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    • pp.227-250
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    • 1999
  • The free radical theory of aging was introduced in 1956 by Denham Harman. This aging theory proposed that normal aging results from random deleterious damage to tissues by free radical and supplying antioxidant lead to decrease oxidative damage, inhibit aging process. In this study, we investigated antioxidantive effects of four Korean constitutional prescriptions for 'Soum' constitution - Palmulgunjatang(Y1), Sipyimiguanjungtang(Y2), Osuyubujayijungtang(Y3) and Seungyangyikkibujatang(Y4). Antioxidative activity of this prescriptions was examined by 1,1-diphenyl-2-picrylhyrdazyl radicals, superoxide anion radicals, peroxyl radical, hydroxyl radical scavenging effects and erythrocyte hemolysis inhibitory effects. Y2 and Y3 were shown to have relatively high antioxidative activity on this methods. In additions, result of the cytoprotective effects of Korean constitutional prescriptions agianst 2,2'-azobis(amidinopropane) dihydrochloride (AAPH), a free radical initiator, induced cytotoxcity in human hepatoblastoma cell line was similarly obtained. On the basis of this result, we assayed the antioxidative effects of Y2 and Y3 on experimental oxidative damage, induced in mouse by 100mg/kg AAPH. Male ICR mouse were given oral administration of 500mg/kg Y2 and Y3 for 4 weeks. Thiobarbuturic acid reactive substance (TBARS) and protein degradation level in liver, plasma and brain as index of oxidative damage were decreased and thiol compound, total antioxidant status in plasma were increased by Y2 administration. But, Y3 injected group was decreased only protein degradation level in brain. Also, glutathione, a potent water-soluble endogenous antioxidant, concentration was increased by Y2 and Y3 administration in liver and brain. However, superoxide dismutase and catalase activity as a major antioxidative enzyme in vivo were not shown change by Y2 and Y3 administration. On the basis of these result, Y2 have an antioxidative effects on both water-soluble fraction and lipid-solube fraction in cell and tissues. But, Y3 has a lower antioxidative effects on lipid-soluble fraction than Y2 in cell and tissues. These results suggest that Y2 has a antioxidative effects by protect the tissue against oxygen free radical mediated oxidative damage and Y3 has a limited antioxidaitve effects on water-soluble fraction in vivo. Therefore, we make report that Y2 is more effective prescriptions for anti-aging or therapeutics of diseases.

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Effect of Obesity and Diabetes on Alzheimer's APP Gene Expression in Mouse Adipose Tissues (비만 및 당뇨가 생쥐 지방조직에서의 Alzheimer's APP 유전자 발현에 미치는 영향)

  • Kim, Jin-Woo;Lee, Yong-Ho
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1012-1018
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    • 2010
  • The aim of this study was to determine whether Alzheimer's amyloid precursor protein (APP) is dysregulated in adipose tissues of C57BL/6 male mice by high-fat diet (HFD) induced obesity, aging, or streptozotocin (STZ)-induced diabetes. APP mRNA expression was examined by quantitative real-time PCR (QPCR) in subcutaneous (SAT) and epididymal adipose tissues (EAT) from mice in 8 different condition groups. By combining conditions of age (16 weeks/26 weeks of age), diet (normal diet (ND)/high-fat diet), and induction of diabetes (non-diabetic/diabetic), 88 mice were divided into 8 different groups. QPCR demonstrated that APP expression in SAT was significantly increased by about two-fold in HFD-induced obese mice compared to both 16 week-old and 26 week-old mice in the ND group (16 weeks p=0.001; 26 weeks p<0.0001), but no changes in EAT was found. Particular effects of aging on APP gene expression were not observed in either adipose tissue depots. Significantly decreased APP expression was found in SAT in STZ-induced diabetic mice fed on ND or HFD at 16 weeks of age (ND p<0.05; HFD p<0.01). Linear regression analysis demonstrated that APP expression levels correlated with body weight in both the non-diabetic group (R=0.657, p<0.0001, n=39) and the diabetic group (R=0.508, p=<0.0001, n=49), but did not correlate with plasma glucose levels, which suggests that decreased APP expression in STZ-induced diabetic mice is most likely due to weight loss rather than hyperglycemia. These data confirm APP dysregulation by weight changes in humans and suggest a possible role linking midlife obesity with the later development of amyloidogenesis in the brain of older patients with Alzheimer's disease.

Gene Expression Profiles in Cervical Cancer with Radiation Therapy Alone and Chemo-radiation Therapy (자궁경부암의 방사선치료 및 방사선항암화학 병용치료에 따른 유전자발현 조절양상)

  • Lee Kyu Chan;Kim Meyoung-kon;Kim Jooyoung;Hwang You Jin;Choi Myung Sun;Kim Chul Yong
    • Radiation Oncology Journal
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    • v.21 no.1
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    • pp.54-65
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    • 2003
  • Purpose : To analyze the gene expression Profiles of uterine ceulcal cancer, and its variation after radiation therapy, with or without concurrent chemotherapy, using a CDNA microarray. Materials and Methods :Sixteen patients, 8 with squamous ceil carcinomas of the uterine cervix, who were treated with radiation alone, and the other 8 treated w14h concurrent chemo-radiation, were Included in the study. Before the starling of the treatment, tumor biopsies were carried out, and the second time biopsies were peformed after a radiation dose of 16.2$\~$27 Gy. Three normal cervix tissues were used as a control group. The microarray experiments were peformed with 5 groups of the total RNAs extracted individually and then admixed as control, pre-radiation therapy alone, during-radiation therapy alone, pre-chemoradiation therapy, and during-chemoradlation therapy. The 33P-iabeled CDNAS were synthesized from the total RNAs of each group, by reverse transcription, and then they were hybridized to the CDNA microarray membrane. The gene expression of each microarrays was captured by the intensity of each spot produced by the radioactive isotopes. The pixels per spot were counted with an Arrayguage, and were exported to Microsoft Excel The data were normalized by the Z transformation, and the comparisons were peformed on the Z-ratio values calculated. Results : The expressions of 15 genes, including integrin linked kinase (ILK), CDC28 protein kinase 2, Spry 2, and ERK 3, were increased with the Z-ratio values of over 2.0 for the cervix cancer tissues compared to those for the normal controls. Those genes were involved In cell growth and proliferation, cell cycle control, or signal transduction. The expressions of the other 6 genes, Including G protein coupled receptor kinase 5, were decreased with the Z-ratio values of below -2.0. After the radiation thorapy, most of the genes, with a previously Increase expressions, represented the decreased expression profiles, and the genes, with the Z-ratio values of over 2.0, were cyclic nucleotlde gated channel and 3 Expressed sequence tags (EST). In the concurrent chemo-radiation group, the genes involved in cell growth and proliferation, cell cycle control, and signal transduction were shown to have increased expressions compared to the radiation therapy alone group. The expressions of genes involved in anglogenesis (angiopoietln-2), immune reactions (formyl peptide receptor-iike 1), and DNA repair (CAMP phosphodiesterase) were increased, however, the expression of gene involved In apoptosls (death associated protein kinase) was decreased. Conclusion : The different kinds of genes involved in the development and progression of cervical cancer were identified with the CDNA microarray, and the proposed theory is that the proliferation signal stalls with ILK, and is amplified with Spry 2 and MAPK signaling, and the cellular mitoses are Increased with the increased expression oi Cdc 2 and cell division kinases. After the radiation therapy, the expression profiles demonstrated 4he evidence of the decreased cancer cell proliferation. There was no sigificant difference in the morphological findings of cell death between the radiation therapy aione and the chemo-radiation groups In the second time biopsy specimen, however, the gene expression profiles were markedly different, and the mechanism at the molecular level needs further study.

Effects of Paeonia Radix Rubra aqua-acupuncture on Adjuvant Arthritis in rats (적작약약침(赤芍藥藥鍼)이 Rat의 Adjuvant 관절염(關節炎)에 미치는 영향(影響))

  • Park, Hee-Soo;Sohn, Seong-Hyoun
    • Journal of Acupuncture Research
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    • v.17 no.1
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    • pp.153-168
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    • 2000
  • To investigate effects of Paeonia Radix Rubra aqua-acupuncture on Adjuvant Athritis in rats, the edema rate, the number of WBC, the quantity of total protein, albumin and globuline in the blood serum were measured and histological test of the muscular tissue were done in the arthritis part. 1. After arthritis of Sprague dawley(SD) rats was induced by injecting Freund's complete adjuvant for 2 weeks, normal saline was injected for the Control group and Paeonia Radix Rubra acua-acupuncture was injected for the Exp. group during 30 days. Selected point was $D{\acute{u}}bf(ST_{35})$ in both groups. And then the inframmator volume of plantar were checked. The volume of the plantar was $0.84{\pm}0.14mm$ in Control group and $0.57{\pm}0.09mm$ in Exp. group, the swelling of plantar was restricted significanily in Exp, group(P<0.05). 2. The number of WBC was $10.34{\pm}0.14(10^3/ml)$ in normal group and $37.47{\pm}5.46(10^3/ml)$ in Control group. It was $25.21{\pm}3.45(10^3/ml)$ in Exp. group. This fact showed that Exp group. with Paeonia Radix Rubra acua-acupuncture was more effective than Control group in the treatment of arthritis(P<0.05). 3. The content of total protein in the blood serum were $6.14{\pm}0.43g/d{\ell}$ in normal group, $7.95{\pm}0.94g/d{\ell}$ in Control group, and $6.82{\pm}1.23g/d{\ell}$ in Exp. group. There was no significance in total protein between Exp. group and Control group from the statistical analysis. 4. The contents of albumin in the blood serum were $2.94{\pm}0.13g/d{\ell}$ in normal group, $2.01{\pm}0.48g/d{\ell}$ in Control group and $2.75{\pm}0.14g/d{\ell}$ in Exp, group. This fact showed that Exp. group had significant increase in the serum albumin from the statistical analysis, in comparison with Control group(P<0.05). 5. The contents of globulin in the blood serum were $3.19{\pm}0.48g/d{\ell}$ in normal group, $4.70{\pm}1.26g/d{\ell}$ in Control group and $3.58{\pm}0.73g/d{\ell}$ in Exp. group. There was no significance in the serum globulin between Exp. group and Control group from the statistical analysis. 6. In histological examinadon, because inflammatory reaction was active, remarkably irregular tissue and large amount of inflammatory cells were found in Control. group. But Exp. group showed that tissues were in some degree recovered, the number of inflamed cells was decreased and inflammation was restrained. From these results, it is shown Paeonia Radix Rubra aqua-acupuncture restrained inflammatory reaction and muscular tissue necrosis in SD rat's plantar was restrained by Freund's complete adjuvant.

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Experimental Study of GuizhiShaoyaoZhimu-Tang on the Rheumatic Pathologic Model Induced by Ajuvant in Rats (계지작약지모탕(桂枝芍藥知母湯)이 Ajuvant 투여로 유발된 풍습성(風濕性) 동물병태모델에 미치는 실험적 연구)

  • Jeong, Hyun-Woo
    • Herbal Formula Science
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    • v.20 no.1
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    • pp.25-40
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    • 2012
  • The purpose of this study explain the experimental effects of Guizhishaoyaozhimu-Tang (桂枝芍藥知母湯) that have clinical efficacy in rheumatoid Arthritis. Materials of present study were Guizhishaoyaozhimu-Tang freeze dried powder (GSZT), Sprague-dawley rats (300 g or so, male), various kinds of needing experimental studis. In order to study the therapeutic effects of GSZT, this Prescription (GSZT 500 mg/kg, 1,000 mg/kg) were administered per oral to the rats with the arthritis induced by Freund's complete adjuvant ($0.2m{\ell}/kg$), several experimental items were measured and compared each other ; that is body weight, rate of edema, analgesic effect by hot plate method, WBC, total protein, TNF-${\alpha}$, IL-10 and expression and localization of H&E, COX-2 staining in synovial tissues from rat with rheumatoid arthritis by immunohistochemical staining using polyclonal COX-2 antibodies. Rats were divided into four groups. Normal group was not treated with Freund's complete adjuvant and treated with DDW 1.0 $m{\ell}$, Control group was treated with Freund's complete adjuvant 0.2 $m{\ell}/kg$ and DDW 1.0 $m{\ell}$, Sample A group was treated with Freund's complete adjuvant 0.2 $m{\ell}/kg$ and GSZT (500 $mg/m{\ell}$) $1.0m{\ell}$, Sample B group was treated with Freund's complete adjuvant 0.2 $m{\ell}/kg$ and GSZT (1,000 $mg/m{\ell}$) 1.0 $m{\ell}$. 0 was day that did not start experiment, 14 was day that confirmed rheumatism induced by Freund's complete adjuvant, 28 was day that completed experiment. The following results were obtained in this study ; Sample A group was increased in body weight, escape time and paw licking time statistical significance compared with Control group, and were decreased in edema, WBC, total protein, TNF-${\alpha}$ with statistical significance compared with Control group. Sample B group was increased in escape time with statistical significance compared with Control group, and were decreased in edema, WBC, total protein with statistical significance compared with Control group. Sample A and Sample B groups were increased in IL-10 compared with Control group, and Sample B group was decreased in TNF-${\alpha}$ compared with Control group. And, Control, Sample A and Sample B groups were showed considerable reduction of positive expression in comparison to Normal group. Especially, Sample B group was most significantly reduction of positive expression than the other groups. From above results, I suggest that GuizhiShaoyaoZhimu-Tang can be used for curing rheumatoid arthritis.

Dlx3 Plays a Role as a Positive Regulator of Osteoclast Differentiation

  • Cha, Ji-Hun;Ryoo, Hyun-Mo;Woo, Kyung-Mi;Kim, Gwan-Shik;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • v.32 no.3
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    • pp.85-91
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    • 2007
  • Dlx3 is a homeodomain protein and is known to playa role in development and differentiation of many tissues. Deletion of four base pairs in DLX3 (NT3198) is causally related to tricho-dento-osseous (TDO) syndrome (OMIM # 190320), a genetic disorder manifested by taurodontism, hair abnormalities, and increased bone density in the cranium. Although the observed defects of TDO syndrome involves bone, little is known about the role of Dlx3 in bone remodeling process. In this study, we examined the effect of wild type DLX3 (wtDlx3) expression on osteoclast differentiation and compared it with that of 4-BP DEL DLX3 (TDO mtDlx3). To examine whether Dlx3 is expressed during RANKL-induced osteoclast differentiation, RAW264.7 cells were cultured in the presence of receptor activator of nuclear factor-B ligand (RANKL). Dlx3 protein level increased slightly after RANKL treatment for 1 day and peaked when the fusion of prefusion osteoclasts actively progressed. When wtDlx3 and TDO mtDlx3 were overexpressed in RAW264.7 cells, they enhanced RANKL-induced osteoclastogenesis and the expression of osteoclast differentiation marker genes such as calcitonin receptor, vitronectin receptor and cathepsin K. Since osteoclast differentiation is critically regulated by the balance between RANKL and osteoprotegerin (OPG), we examined the effect of Dlx3 overexpression on expression of RANKL and OPG in C2C12 cells in the presence of bone morphogenetic protein 2. Overexpression of wtDlx3 enhanced RANKL mRNA expression while slightly suppressed OPG expression. However, TDO mtDlx3 did not exert significant effects. This result suggests that inability of TDO mtDlx3 to regulate expression of RANKL and OPG may contribute to increased bone density in TDO syndrome patients. Taken together, it is suggested that Dlx3 playa role as a positive regulator of osteoclast differentiation via up-regulation of osteoclast differentiation-associated genes in osteoclasts, as well as via increasing the ratio of RANKL to OPG in osteoblastic cells.

Electrophoresis of Proteins in the Tissues from Witches' - Broom infected Jujube Tree (빗자루병(病)에 감염(感染)된 대추나무 조직내(組織內) 단백질(蛋白質)의 전기영동(電氣泳動))

  • La, Yong Joon;Park, Weon Chul;Park, Weon Mook;Lee, Yong Se
    • Journal of Korean Society of Forest Science
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    • v.66 no.1
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    • pp.79-81
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    • 1984
  • Protein profiles of healthy and witches' broom (mycoplasma) infected jujube plants (Zizyphus jujuba) were investigated by 2-30% linear gradient polyacrylamide gel electrophoresis. Distinct differences in band patterns between healthy and infected samples were observed. Gels from samples of healthy leaves showed a characteristic protein band in the 50kd-range, which was not detected in infected leaves. Band with 25kd was more distinct in the infected leaves, whereas band with 198kd was more apparent in the healthy leaves. Healthy-looking leaves in the infected samples demonstrated the characteristic band of 50kd with less intensity showing intermediate pattern between healthy leaves and infected leaves. In contrast with leaf samples, gels from infected stem samples showed a characteristic band in the 335kd-range which was absent in healthy stem samples.

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Immunocytochemical Localization of Metallothionein in Gastric Adenocarcinoma (위암 조직내 Metallothionein의 면역 세포화학적 연구)

  • Yang, Seung-Ha;Shin, Kil-Sang;Kim, Wan-Jong
    • Applied Microscopy
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    • v.32 no.4
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    • pp.411-419
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    • 2002
  • Metallothionein (MT) is a family of ubiquitous, low molecular weight ($6,000{\sim}7,000D$), cysteine-rich ($30{\sim}35%$) inducible protein with a high affinity to metal ions and has no aromatic amino acids and histidine. Some of the known functions of MT include detoxification of heavy metals and alkylating agents and neutralization of free radicals. Also, this protein has been reported to involve in tumor pathophysiology and therapy resistance. MT expression may affect a number of cellular processes including gene expression, apoptosis, proliferation and differentiation. Many reports on the physiological and biochemical properties of MT have been published, but ultrastructural reports on the localization of MT in human gastric cancer tissues are extremely rare. The present study was undertaken to examine the ultrastructural features and the localization of MT within the gastric adenocarcinoma. Ultrastructures of gastric cancer cells were characterized by the high nuclear cytoplasmic ratio, the interdigitation between cells, the irregular nucleus containing much heterochromatin and the wide distribution of free ribosomes in the cytoplasm. Immunohistochemical reaction for MT was prominent in the gastric adenocarcinoma. And the immunogold labellings were more prominent within the nucleus than the cytoplasm. Particularly, immunogold particles were numerously seen at nulcleolus or nucleolar associated heterochromatin. These results suggest that MT expression by gastric cancer cells is associated with cell proliferative activity and is possibly synthesized in the cytoplasm, and then the protein is transported into the nucleus to participate in any transcriptional steps.

RalA-binding Protein 1 is an Important Regulator of Tumor Angiogenesis (Tumor angiogenesis에 있어서 RLIP76의 중요성)

  • Lee, Seunghyung
    • Journal of Life Science
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    • v.24 no.5
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    • pp.588-593
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    • 2014
  • Tumor angiogenesis is important in tumorigenesis and therapeutic interventions in cancer. To know inhibitor and effector of tumor angiogenesis in cancer, the specific gene of tumor and angiogenesis may develop the mechanisms of cancer suppression and therapy. Recently, we described the role of RalA-binding protein 1 (RLIP76) in tumor angiogenesis. Tumor vascular volumes were diminished, and vessels were fewer in number, shorter, and narrower in RLIP76 knockout mice than in wild-type mice. Moreover, angiogenesis in basement membrane matrix plugs was blunted in the knockout mice in the absence of tumor cells, with endothelial cells isolated from the lungs of these animals exhibiting defects in migration, proliferation, and cord formation in vitro. RLIP76 is expressed in most human tissues and is overexpressed in many tumor types. In addition, the protein regulates tumorigenesis and angiogenesis in vivo and in vitro. As the export of chemotherapy agents is a prominent cellular function of RLIP76, it is a major factor in mechanisms of drug resistance. Moreover, RLIP76 acts as a selective effector of the small GTPase, R-Ras, and regulates R-Ras signaling, leading to cell spreading and migration. Furthermore, in skin carcinogenesis, RLIP76 knockout mice are resistant, with tumors that form showing diminished angiogenesis. Thus, RLIP76 is required for efficient endothelial cell function and angiogenesis in solid tumors.

Characterization of a non-specific Lipid Transfer Protein (ns-LTP) promoter from poplar (Populus alba × P. glandulosa) (현사시나무(Populus alba × P. glandulosa)에서 분리한 non-specific Lipid Transfer Protein (ns-LTP) 프로모터의 특성 분석)

  • Cho, Jin-Seong;Noh, Seol Ah;Choi, Young-Im
    • Journal of Plant Biotechnology
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    • v.42 no.4
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    • pp.356-363
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    • 2015
  • In order to study genetic engineering in trees, the characterization of genes and promoters from trees is necessary. We isolated the promoter region (867 bp) of Pagns-LTP from poplar (P. alba ${\times}$ P. glandulosa) and characterized its activity in transgenic poplar plants using a ${\beta}$-glucuronidase (GUS) reporter gene. High-level expression of the Pagns-LTP transcript was found in poplar roots, while comparatively low-level expression was found in the young leaves. Pagns-LTP mRNA was not detected in other poplar tissues. Additionally, transgenic poplar plants that contained a Pagns-LTP promoter fused to a GUS reporter gene, displayed tissue-specific GUS enzyme activity localized in root tissue. In silico analysis of the Pagns-LTP promoter sequence reveals the presence of several cis-regulatory elements responsive to phytohormones, biotic and abiotic stresses, as well as those regulating tissue-specific expression. These results demonstrate that the Pagns-LTP promoter has tissue-specific expression activity in poplar roots and leaves that may be involved in organ development and plant resistance to various stresses. Therefore, we anticipate that the Pagns-LTP promoter would be a useful tool to genetically optimize woody plants for functional genomics.