• Title/Summary/Keyword: marker substance

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Development and Research into Functional Foods from Hydrolyzed Whey Protein Powder with Sialic Acid as Its Index Component - III. Bacterial Reverse Mutation Testing of Hydrolyzed Whey Protein Powder Containing Normal Concentration of Sialic Acid (7%) with Enzyme Separation Method - (Sialic Acid를 지표성분으로 하는 유청가수분해단백분말의 기능성식품 개발연구 - III. 효소분리로 7% Siailc Acid가 표준적으로 함유된 유청가수분해단백분말의 미생물복귀돌연변이시험 연구 -)

  • Kim, Hee-Kyong;Noh, Hye-Ji;Cho, Hyang-Hyun;Koh, Hong Bum
    • Journal of Dairy Science and Biotechnology
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    • v.34 no.2
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    • pp.137-144
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    • 2016
  • The ultimate research goal of the current study was a development of hydrolyzed whey protein powder (7%-GNANA) manufactured with normal content of sialic acid, a marker compound, that is naturally occurring at 7% concentration in GMP obtained from the milk protein. GMP is a safe food, used worldwide in infant and baby foods, etc. The test substance was prepared using (7% sialic acid containing) GMP as a raw material, and then using alcalase, an enzyme approved as a food additive, after separation of sialic acid with 100% efficiency and 7%-GNANA (containing 7% sialic acid and protein; product name: HELICOBACTROL-7) provided by MEDINUTROL Inc. (Korea). Bacterial reverse mutation (Ames) test was conducted in accordance with GLP Guideline using the test substance specified above. To identify its mutagenic potential against microorganisms, histidine auxotrophic strains of Salmonella Typhimurium, TA98, TA100, TA1535, and TA1537, and tryptophan auxotrophic strain of Escherichia coli, WP2uvrA, were used. The bacterial reverse mutation (Ames) test was performed by dividing the test substances into five different concentration groups (0, 61.7, 185, 556, 1,670, $5,000{\mu}g/plate$). Results of this experiment did not reveal repetitive increase of colony generating values or positive criteria for reverse mutagenicity for any concentration of test substances in any of the five strains, regardless of the presence of a metabolic activation system, and no dose-dependency was identified. In conclusion, the safety of 7%-GNANA test substance was verified by bacterial reverse mutation test conducted before registration of 7%-GNANA as a food additive.

Bacterial Reverse Mutation Test Evaluation of Hydrolyzed GMP Powder Containing Highly Concentrated Sialic Acid (23%) produced by Enzyme Separation and Solvent Enrichment Method (효소분리 및 용매정제법으로 제조한 고농도 Sialic Acid(23%)가 함유된 GMP 가수분해분말의 미생물복귀돌연변이시험 연구)

  • Kim, Hee-Kyong;Cho, Hyang-Hyun;Noh, Hye-Ji
    • Journal of Dairy Science and Biotechnology
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    • v.34 no.2
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    • pp.91-98
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    • 2016
  • The goal of this study was to develop hydrolyzed whey protein powder (23%-GNANA) manufactured with high content of sialic acid, a marker compound that is usually present at 7% concentration in GMP obtained from the milk protein. It is a safe food, used worldwide in infant and baby foods, etc. The test substance was prepared using (7% sialic acid containing) GMP as a raw material. Alcalase, an enzyme approved as a food additive, was used after separating sialic acid, with 100% efficiency, and 23%-GNANA (composed of 23% sialic acid and protein; product name: HELICOBACTROL-23), provided by MEDINUTROL Inc. (Korea), manufactured to have high (23%) content through ethanol soaking and enrichment. Bacterial reverse mutation (Ames) test was conducted in accordance with the GLP Guideline using the test substance specified above. To detect its mutagenicity potential in microorganisms, histidine auxotrophic strains of Salmonella typhimurium, TA98, TA100, TA1535, and TA1537, and tryptophan auxotrophic Escherichia coli strain, WP2uvrA, were used. The bacterial reverse mutation (Ames) test was performed using five concentrations of the test substances (0, 61.7, 185, 556, 1,670, $5,000{\mu}g/plate$). The evaluation did not reveal repetitive increase of colony generating values and positive criteria for reverse mutagenicity for any tested concentration in the five strains regardless of the presence of metabolic activation system, and no dose-dependency. In conclusion, the safety of 23%-GNANA test substance was verified by the bacterial reverse mutation test conducted before registration of 23%-GNANA as a food additive.

Genetic Diversity and Metabolite Analysis of Gastrodia elata by Inter-Simple Sequence Repeats (ISSR) Markers (ISSR 표지에 의한 천마의 유전 다양성분석 및 기능성 물질분석)

  • Kim, Hyun Tae;Kim, Ji Ah;Park, Eung Jun
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.6
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    • pp.440-446
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    • 2012
  • Gastrodia elata, an achlorophyllous orchid plant, is rare medicinal plant. We investigated the genetic diversity in G. elata from 4 locations by using Inter-Simple Sequence Repeats (ISSR) markers. Shannon's information Index (S.I.) indicating genetic diversity ranged from 0.255 (Pocheon) to 0.322 (Muju) with the mean of 0.29. The level of genetic diversity was lower than other plant and most genetic diversity was allocated among individuals within populations (26.81%). The UPGMA dendrogram based on genetic distance failed in showing decisive geographic relationship. In the case of gastrodin (GA), the major components in G. elata, Sangju was highest. The ergothionine (ERG) was detected a lot of contents in Muju and Pocheon. In conclusion, our results is very important information for explaining relationship of genetic variation and functional substances without the effects of environment factors and developing genetic marker by ISSR in G. elata, which may be responsible for the development of breeds with a lot of functional substance in G. elata.

Inhibitory effects of fenbendazole, an anthelmintics, on lipopolysaccharide-activated mouse bone marrow cells (지질다당류로 활성화된 마우스 골수세포에서 구충제 Fenbendazole의 억제 효과)

  • Park, Seo-Ro;Joo, Hong-Gu
    • Korean Journal of Veterinary Research
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    • v.61 no.3
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    • pp.22.1-22.7
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    • 2021
  • Fenbendazole (FBZ) is a commonly used anthelmintics in veterinary medicine that has recently been found to have anticancer effects in humans. On the other hand, few studies have examined the anti-inflammatory effects of FBZ, and its mechanism is unknown. In this study, mouse bone marrow cells (BMs) were treated with lipopolysaccharide (LPS), a representative inflammation-inducing substance, to generate a situation similar to osteomyelitis in vitro. The effect of FBZ on inflammatory BMs was examined by measuring the metabolic activity, surface marker expression, cell nuclear morphology, and mitochondrial membrane potential (MMP) of BMs. FBZ decreased the metabolic activity and MMP of LPS-treated BMs. Annexin V-fluorescein isothiocyanate/propidium iodide staining and Hoechst 33342 staining showed that FBZ reduced the number of viable cells and induced the cell death of inflammatory BMs. In addition, FBZ reduced the proportion of granulocytes more than B lymphocytes in LPS-treated BMs. Overall, FBZ induces cell death by destabilizing the MMP of LPS-induced inflammatory BMs. In addition to anthelmintic and anticancer agent, FBZ can play a role as an anti-inflammatory agent.

Effects of FK224, a $NK_1$ and $NK_2$ Receptor Antagonist, on Plasma Extravasation of Neurogenic Inflammation in Rat Airways (미주 신경의 전기적 자극으로 유발된 백서의 기도내 혈장 유출에 대한 FK224의 효과)

  • Shim, Jae-Jeong;Lee, Sang-Yeub;Lee, Sang-Hwa;Park, Sang-Myun;Seo, Jeong-Kyung;Cho, Jae-Yun;In, Kwang-Ho;Yoo, Se-Hwa;Kang, Kyung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.5
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    • pp.744-751
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    • 1995
  • Background: Asthma is an inflammatory disease because there are many inflammatory changes in the asthmatic airways. Axon reflex mechanisms may be involved in the pathogenesis of asthma. Sensory neuropeptides are involved in this inflammation, which is defined as neurogenic inflammation. Substance p, neurokinin A, and neurokinin B may be main neuropeptides of neurogenic inflammation in airways. These tachykinins act on neurokinin receptors. Three types of neurokinin receptors, such as $NK_1$, $NK_2$, and $NK_3$, are currently recognized, at which substance p, neurokinin A, and neurokinin B may be the most relevant natural agonist of neurogenic inflammation in airways. The receptor subtypes present in several tissues have been characterized on the basis of differential sensitivity to substance p, neurokinin A, and neurokinin B. Plasma extravasation and vasodilation are induced by substance p more potently than by neurokinin A, indicating NK1 receptors on endothelial cells mediate the response. But airway contraction is induced by neurokinin A more potently than by substance P, indicating the $NK_2$ receptors in airway smooth muscles. These receptors are used to evaluate the pathogenesis of brochial asthma. FK224 was identified from the fermentation products of Streptomyces violaceoniger. FK224 is a dual antagonist of both $NK_1$ and $NK_2$ receptors. Purpose: For a study of pathogenesis of bronchial asthma, the effect of FK224 on plasma extravasation induced by vagal NANC electrical stimulation was evaluated in rat airway. Method: Male Sprague-Dawley rats weighing 180~450gm were anesthetized by i.p. injection of urethane. Plasma extravasation was induced by electrical stimulation of cervical vagus NANC nerves with 5Hz, 1mA, and 5V for 2 minutes(NANC2 group) and for sham operation without nerve stimulation(control group). To evaluate the effect of FK224 on plasma extravasation in neurogenic inflammation, FK224(1mg/kg, Fujisawa Pharmaceutical Co., dissolved in dimethylsulphoxide; DMSO, Sigma Co.) was injected 1 min before nerve stimulation(FK224 group). To assess plasma exudation, Evans blue dye(20mg/kg, dissolved in saline) was used as a plasma marker and was injected before nerve stimulation. After removal of intravascular dye, the evans blue dye in the tissue was extracted in formamide($37^{\circ}C$, 24h) and quantified spectrophotometrically by measuring dye absorbance at 629nm wavelength. Tissue dye content was expressed as ng of dye per mg of wet weight tissue. The amount of plasma extravasation was measured on the part of airways in each groups. Results: 1) Vagus nerve(NANC) stimulation significantly increased plasma leakage in trachea, main bronchus, and peripheral bronchus compared with control group, $14.1{\pm}1.6$ to $49.7{\pm}2.5$, $17.5{\pm}2.0$ to $38.7{\pm}2.8$, and $12.7{\pm}2.2$ to $19.1{\pm}1.6ng$ of dye per mg of tissue(mean ${\pm}$ SE), respectively(p<0.05). But there was not significantly changed in lung parenchyma(p>0.05) 2) FK224 had significant inhibitory effect upon vagal nerve stimulation-induced airway plasma leakage in any airway tissues of rat,such as trachea, main bronchus, and peripheral bronchus compared with vagus nerve stimulation group, 49%, 58%, and 70%, respectively(p<0.05). Inhibitory effect of FK224 on airway plasma leakage in neurogenic inflammation was revealed the more significant in peripheral bronchus, but no significant in lung parenchyma. Conclusion: These results suggest that FK224 is a selective NK receptor antagonist which effectively inhibits airway plasma leakage induced by the endogenous neurotransmitters relased by neurogenic inflammation in rat airway. Tachykinin receptor antagonists may be useful in the treatment of brochial asthma.

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The Effect of Seasoning on the Intestinal Absorption -Absorption by Passive Transport and the Effect of Red Pepper- (조미료가 창자 운동과 흡수기능에 미치는 영향 -소장의 피동적 흡수에 대한 고추의 영향-)

  • Shin, Dong-Hoon;Kim, Joong-Soo;Koh, Jae-Pyong;Ahn, Seung-Woon
    • The Korean Journal of Physiology
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    • v.7 no.1
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    • pp.23-31
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    • 1973
  • Numerous factors concern with the absorption of substances through the membrane of the gastrointestinal tract. To simplify the experimental condition, present work has been restricted to observe the disappearance rate of substance from the intestinal loop which was made in the jejunum, 70 cm apart from the pylorus of the adult rabbit. The purpose of the study is to clarify the absorption of urea through the jejunal wall is solely attributable to the concentration difference between the luminal fluid and plasma, and to observe the effect of adding red pepper upon the rate of absorption. The rabbits were anesthetized with nembutal, 35mg/kg I.V. Jejunal loop was made by ligating at 2 spots, 70 cm and 80cm apart from the pylorus. After rinsing with normal saline solution through the polyethylene tubing inserted from the end of the loop, 8 ml of test solution was placed through the same tubing. The test solution contained 200 mg% of urea and 150mg% of polyethylene glycol(M.W. 4,000) in normal saline solution. Right after placing the test solution the first specimen was taken through the tubing, and successive samplings were performed at 5, 10, 20, and 30 minutes. Logarithm of the difference of urea concentration between the luminal fluid and plasma was plotted against time elapsed after the onset of the experiment. If straight line is revealed, it would verify the nature of transport mechanism as diffusion, obeying the Fick's principle. The concentration of polyethylene glycol (PEG) was also measured in order to examine the change in the volume. PEG was used as the marker substance because it is not absorbable in the intestinal tract. Consequently the concentration of PEG relates inversely to the volume of the loop. Instantaneous concentration of urea in the loop times the volume will give the amount of urea remaining in the luminal fluid. The change in the amount of any substance is directly relate to the volume of the compartment and differs from the change in the concentration which is independent of the volume. After completion of the experiment without red pepper, it was added in the test solution and was centrifuged after thorough mixing. Supernatant of the mixture was placed in the loop and similar sampling were performed with the same time intervals that of previous run in order to observe the effects of the red pepper on the passive transport of the water soluble small substance, urea. The results obtained were as follows: 1. Logarithm of the concentration difference of urea between the luminal fluid and plasma was diminished exponentially as time elapsed. The decay constant in the experiment without red pepper was 0.0563/min. By adding red pepper in the test solution as much as the concentration rose to 4,000 mg% and 8,000 mg%, the decay constants were lowered to 0.0493/min and to 0.0506/min, respectively. The time interval by which the concentration difference dropped to one half of the initial value was prolonged. Without red pepper the half concentration time was 13.30 minutes, and by adding extract of red pepper, 15.31 minutes and 15.71 minutes were revealed. 2. The profile of the diminishing rate of tile amount of urea was quite different from that of the concentration because of the change in the volume of the loop during the observed period. 3. By adding the extract of red pepper, it slowed down the rate of absorption of urea in the intestinal loop, suggesting an increase in the diffusional barrier. 4. Larger dosage of red pepper brought an increase in the secretion of intestinal fluid with concomitant expansion of the luminal volume, and the retardation of the absorption of urea was noticed. This effect was largely dependent on the sensitivity of the individual animal to the red pepper, extract. The amount of urea remained after 10 minutes interval was 55.5% of the initial amount in the experiment without red pepper. On the other hand it was not consistent after administration of red pepper, showing 50.6% and 66.5% of the initial figures by adding 400 mg and 800 mg of red pepper in the test solution, respectively. It was postulated that symptom of diarrhea often encountered by taking a hot (red pepper) food might be attributable to the increase of secretion and the retardation of absorption in the intestinal tract.

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Changes of Phytohormones in Rice and Tobacco Protoplasts under Different Cultural Conditions (벼와 담배의 원형질체 배양조건에 따른 식물호르몬의 변화)

  • 황태익;민경수;임현옥;안장순
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.1
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    • pp.33-41
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    • 1997
  • This study was carried out to establish simple and easy methods to judge the survival, senescence and death of the protoplasts in culture system by identifying the marker substance related to metabolic status of the cells. When rice and tobacco protoplasts were cultured in MS and KM-8P media containing 2,4-D or coconut milk ABA decreased especially in the media containing coconut milk, but GA$_3$, IAA and zeatin increased as the cultures progressed. The decrease of ABA and increase of zeatin was especially remarkable. When the supraoptimal amount of osmoticum (mannitol) was added to the culture media ABA decreased after a momentary increase, but other growth hormones slowly increased as the concentration of the osmoticum increased. Contents of individual hormones were contrasted when protoplasts rice and tobacco were cultured on the same medium containing 10mM super mine or NaCl. Tobacco protoplasts were more sensitive to NaCl stress and stopped protoplast division at the late stage of culture. Protoplast viability decreased greatly in 48 hours when the protoplast were at 32$^{\circ}C$ on a medium lacking several components. ABA content increased up to 10 days from incubation in negative proportion to the protoplast viability. On the other hand contents of other growth hormones, especially zeatin, decreased. The present results clearly showed that the contents of individual growth hormones in the plant protoplasts in culture varied sensitively in response to environmental factors that they are faced with. This indicates that the physiological states of the protoplast, such as survival, senescence or death can be simply judged based on the quantitative analysis of those hormones by ELISA.

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Hepatocyte protection and antioxidant effect of Citri Unshius Pericarpium against cadmium-induced oxidative stress (카드뮴으로 유발된 산화 스트레스에 대한 진피의 간세포 보호 및 항산화 효과)

  • Noh, Gyu Pyo;Byun, Sung Hui;Jung, Dae Hwa;Lee, Jong Rok;Park, Sook Jahr;Kim, Sang Chan
    • Herbal Formula Science
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    • v.28 no.4
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    • pp.327-337
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    • 2020
  • Objective : Citri Unshius Pericarpium is the dried peel of mature fruit of Citrus unshiu Markovich and has been used to treat indigestion, vomiting, and removal of phlegm. This study investigated the hepatoprotective and antioxidant effect of CEE (Ethanol extract of Citri Unshius Pericarpium) in cadmium (CdCl2)-treated HepG2 cells. Methods : Component analysis of Citri Unshius Pericarpium was analyzed by UPLC with C18 column. Cell viability was determined by MTT assay. The enzyme activity of superoxide dismutase (SOD) and the level of reactive oxygen species (ROS) and reduced glutathione (GSH) were analyzed using commercially available kits. Results : Cadmium caused severe HepG2 cell death. Cadmium also increased ROS production, consistent with depletion of GSH and inhibition of the SOD enzyme. However, CEE treatment reduced cell death and relieved oxidative stress caused by cadmium toxicity. CEE lowered ROS levels and improved depletion of GSH levels. CEE also enhanced the enzymatic activity of SOD. In component analysis, hesperidin was the most abundant of the five marker compounds (Narigenin, Narigin, Narirutin, Hesperidin and Hesperidin), which assumes that hesperidin partially contributed to the antioxidant activity of CEE. Conclusion : These results suggested that CEE could be a potential substance to solve heavy metal-related health problems. In particular, inhibition of oxidative stress by CEE can be a way to treat liver damage caused by cadmium.

Validation and Determination of Glycyrrhizic Acid as a Marker Substance in Bu-Zhong-Yi-Qi-Tang by HPLC/DAD (HPLC/DAD에 의한 보중익기탕 중의 글리시리진산 정량분석법의 확립)

  • Baek, Ju-Hyun;Kim, Sun-Min;Ahn, Ji-Won;Cho, Chang-Hee;Oh, Mi-Hyune;Cho, Jung-Hee;Lee, Mi-Kyeong;Kim, Hyo-Jin
    • YAKHAK HOEJI
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    • v.52 no.1
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    • pp.7-11
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    • 2008
  • A high performance liquid chromatographic (HPLC) method for the determination of glycyrrhizic acid was developed for the quality control of traditional herbal medicinal preparation Bu-Zhong-Yi-Qi-Tang (BZYQT), which is well-known herbal medicine used as tonic. RP-HPLC analysis was carried out using Capcell pak $C_{18}$ MG column $(5\;{\mu},\;150{\times}4.6\;mm)$ and a mobile phase consisting of acetonitrile and water containing 0.03% phosphoric acid (pH 2.46) at a flow rate of 1.0 ml/min. The optimum wavelength for the detection of the glycyrrhizic acid was found at 250 nm using diode-array UV/VIS detector. The glycyrrhizic acid in BZYQT shows good linearity $(r^2>0.999)$ in the range of $15\;{\mu}g/ml$ to 500 ${\mu}g/ml$. The limit of detection (LOD) was less than 5 ng and R.S.D for intra-day and inter-day reproducibility was less than 7%. The mean recovery of the glycyrrhizic acid was $97.3{\sim}113.0%$. These results suggest that the developed HPLC method is simple and efficient, and could be contributed for the quality control of commercial BZYQT products.

Migration Activity of Chicken Gonadal Primordial Germ Cells (gPGCs) and Post-transfer Localization of LacZ-transfected gPGCs in the Embryonic Gonads

  • Jeong, D.K.;Han, J.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.9
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    • pp.1227-1231
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    • 2002
  • A powerful tool for chicken transgenesis could be established by employing a germline chimera production through primordial germ cell transplantation. This study was conducted to examine whether foreign gene-transfected gonadal primordial germ cells (gPGCs) have a migration activity into the gonad after transfer to recipient embryos. In Experiment 1, gPGCs of Korean Ogol Chicken were retrieved from 5.5-day-old embryos and subsequently transferred to the dorsal aorta of 2.5-day-old White Leghorn embryos after being labeled with PKH26 fluorescent dye. To confirm migration activity after transplantation, recipient embryos were sacrificed and examined on 3 days after transfer. Sex determination was concomitantly undertaken to examine whether sex of recipient embryos could affect the migration activity of gPGCs. All of embryonic gonads examined showed positive signals with PKH26 fluorescence and W-chromosome specific band by polymerase chain reaction (PCR) was detected in male embryos when gPGCs with ZW chromosome were transferred to recipient embryos. In Experiment 2, retrieved gPGCs were transfected with LacZ gene-containing cytomegalovirus promoter ($pCMV{\beta}$) by electroporation and subsequently transferred to recipient embryos. LacZ gene expression was identified in the gonads of 6 or 10-day-old recipient embryos and hatched-chicks. A total of 20 embryos and 12 hatched-chicks were examined and 11 of them (10 embryos and one hatched chicken; 11/32=34.4%) expressed $\beta$-galactosidase, a marker substance of LacZ gene. The results of this study demonstrated that foreign gene-transfected gPGCs can migrate and settle down into the gonad after being transferred into the blood vessel of the recipient embryos. This established technique will contribute to developing a peer biotechnology for transgenic chicken.