• Title/Summary/Keyword: 형질 세포

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Chromosomal Localization and Mutation Detection of the Porcine APM1 Gene Encoding Adiponectin (Adiponectin을 암호화하는 돼지 APM1 유전자의 염색체상 위치파악과 돌연변이 탐색)

  • Park, E.W.;Kim, J.H.;Seo, B.Y.;Jung, K.C.;Yu, S.L.;Cho, I.C.;Lee, J.G.;Oh, S.J.;Jeon, J.T.;Lee, J.H.
    • Journal of Animal Science and Technology
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    • v.46 no.4
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    • pp.537-546
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    • 2004
  • Adiponectin is adipocyte complement-related protein which is highly specialized to play important roles in metabolic and honnonal processes. This protein, called GBP-28, AdipoQ, and Acrp30, is encoded by the adipose most abundant gene transcript 1 (APM1) which locates on human chromosome 3q27 and mouse chromosome 16. In order to determine chromosomal localization of the porcine APM1, we carried out PCR analysis using somatic cell hybrid panel as well as porcine whole genome radiation hybrid (RH) panel. The result showed that the porcine APM1 located on chromosome 13q41 or 13q46-49. These locations were further investigated with the two point analysis of RH panel, revealed the most significant linked marker (LOD score 20.29) being SIAT1 (8 cRs away), where the fat-related QTL located. From the SSCP analysis of APM1 using 8 pig breeds, two distinct SSCP types were detected from K~ native and Korean wild pigs. The determined sequences in Korean native and Korean wild pigs showed that two nucleotide positions (T672C and C705G) were substituted. The primary sequence of the porcine APM1 has 79 to 87% identity with those of human, mouse, and bovine APM1. The domain structures of the porcine APM1 such as signal sequence, hypervariable region, collagenous region. and globular domain are also similar to those of mammalian genes.

Cytotaxonomic study of Korean Euphorbia L. (Euphorbiaceae) (한국산 대극속(Euphorbia L., Euphorbiaceae)의 세포분류학적 연구)

  • Chung, Gyu Young;Oh, Byoung-Un;Park, Ki-Ryong;Kim, Joo-Hwan;Kim, Mi Suk;Nam, Gi-Heum;Jang, Chang-Gee
    • Korean Journal of Plant Taxonomy
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    • v.33 no.3
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    • pp.279-293
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    • 2003
  • Somatic chromosomes about 13 taxa of Korean Euphorbia L. was investigated to estimate its taxonomic significance. Somatic chromosome numbers of treated taxa were 2n= 12, 20, 22, 28, 40, 42, 56, therefore basic chromosome numbers of those were x=6, 7, 10, 11. The chromosome numbers of E. pallasii Turcz. (2n=20), E. hylonoma Hand.-Mazz (2n=20.), E. fauriei H. L$\acute{e}$v. & Vaniot ex H. L$\acute{e}$v (2n=28) and E. jolkini Boiss. (2n=28) were determined for the first time in this study. The chromosome numbers of four taxa were same as previous ones; E. sieboldiana Moor. & Decne. (2n=20), E. ebracteolata Hayata (2n=20), E. humifusa Willd. ex Schlecht. (2n=22). But those of six taxa were different; E. esula L (2n= 16, 20, 60, 64 vs 2n=20), E. helioscopia L. (2n=12, 42 vs 2n=42), E. lucorum Rupr. (2n=28, 40 vs 2n=56), E. pekinensis Rupr. in Maxim. (2n=24 vs 2n=28, 56), E. maculata L. (2n=28, 42 vs 2n=12), E. supina Raf. (n=7 vs 2n=40). E. ebracteolata, E. pallasii and E. hylonoma were distingushcd from the other taxa by the chromosome numbers, size and satellites, E. maculata, E. humifusa, E. supina had the different basic and somatic chromosome numbers in spite of the similar morphological. anatomical and palynological chracters. The chromosomal character of Korean Euphorbia was supported the Ma and Hu's systems, and as above results, it was found to be a good character in delimiting above sections and estimating relationships for some species.

Effect of Sucrose and Polybrene on the Gene Transfer into Porcine Oocytes using Retroviral Vector (레트로 바이러스 벡터를 이용한 돼지난자에의 유전자 전이에 있어 Sucrose와 Polybrene의 효과)

  • Kim, . K.S.;M.S. Kwon;J.Y. Ju;Kim, K.S.;Kim, T.;Lee, H.T.;K.S. Chung
    • Korean Journal of Animal Reproduction
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    • v.26 no.2
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    • pp.153-163
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    • 2002
  • In vitro matured porcine oocytes have very small volume of perivitellinspace (PVS). In these respect, the effect of sucrose and polybrene on the efficiency of gene transfer was investigated. As a gene (hGH) transfer vehicle, vesicular stomatitis virus glycoprotein pseudotyped retroviral vector (VSV-G) was used. Sucrose treatment has no detrimental effect on the rates of cleavage and resulted in the enlargement of PVS for the efficient introduction of retroviral vector stocks. Introduction rates of retrovirus in 0.5, 1, 2, 3 % sucrose treatment group were higher than that of the non-treatment group (39.3, 43.3, 35.7, 40.7 % vs. 8.3 %), respectively. In addition, we observed that sucrose pretreatment during injection procedure significantly reduce the frequency of polyspermy. In general, polybrene is a polycation essential for retrovirus transduction. The groups with the addition of 0.5, 5, 50$\mu\textrm{g}$/$m\ell$ polybrene exhibited a significant effect on gene transfer compared to that of the non-addition group (56.5, 50.0, 57.1 % vs. 34.6 %), respectively But, when the oocytes were co-injected with retrovirus and 50$\mu\textrm{g}$/$m\ell$ polybrene, the rates of cleavage and blastocyst development were 43.3 and 4.6%, respectively. This rates were lower than those of the non-addition group (70.0 and 17.3 %). In conclusion, sucrose pretreatment have increased efficiency of retroviral mediated gene transfer in porcine oocytes with no damage on in vitro fertilization and embryo development. In addition, sucrose pretreatment was beneficial in polyspermy inhibition. Presence of polybrene during microinjection showed a beneficial effect on the gene transfer in porcine oocytes, in low concentration. And these results will provide an useful tool for production of transgenic pigs by retroviral mediated gene transfer.

Analysis of phylogenetic relationship among Korean Pinellia Tenore (Araceae) using RAPD markers (RAPD markers에 의한 한국산 반하속 식물의 유연관계 분석)

  • Tae, Kyoung-Hwan;Kim, Dong-Kap;Kim, Joo-Hwan
    • Korean Journal of Plant Taxonomy
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    • v.35 no.3
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    • pp.161-174
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    • 2005
  • In order to presume the relationships between two species of P. ternata and P. tripartia, and their populations of the Korean Pinellia, RAPD analysis was performed. The length of the amplified DNA fragments ranged from 300 to 2,500bp. Seventy scorable RAPD makers were found from the PCR reactions with 7 random oligoprimers and were analyzed by Nei-Li's genetic coefficient. Also, some regional groups instead of same taxa were clustered from the phenogram of UPGMA analysis and NJ tree. Populations within each species were clustered at low genetic distance, there had the closed relationship. According to the regional individuals, Pinellia ternata was showed the variation pattern of morphological (leaf shape and flower color) and cytological characters(somatic chromosome numbers). So we suggested to difference of characteristic variety based on variety of habitat. According to this study, new species (Pinellia sp.) was affiliated with Pinellia and had the closest relationship with Hallasan and Japan population. The RAPD data was very useful to define the genetic variation and to discuss the relationships among the intraspecific taxa and their populations of the Korean Pinellia.

Analysis of Populus cpDNA by Restriction Fragment Length Polymorphism(RFLP) Technique (RFLP기법(技法)을 이용(利用)한 포플러 엽록체(葉綠體) DNA의 분석(分析))

  • Lee, J.S.;Noh, E.W.;Lee, S.K.;Kwon, K.W.
    • Journal of Korean Society of Forest Science
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    • v.83 no.1
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    • pp.20-24
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    • 1994
  • In woody species with a long life span, the studies on inheritance of any trait may be very time consuming and laborious. Chloroplast DNA(cpDNA) has been a valuable tool in such studies since it has several unique features such as limited genome size and cytoplasmic inheritance. In the present study, cpDNAs from five different species of Populus(P. alba, P. glandulosa, P. alba${\times}$P. glandulosa, P. davidiana, and P. nigra), and Nicotiana tabacum were compared with regard to restriction fragment length polymophism. The results showed that cpDNAs among the species were very conserved, although some polymorphisms were observed when the DNAs were digested with restriction enzyme EcoRI or KphI. The other enzymes (Bgl II, and PstI) tested produced identical restriction fragmentation pattern among the species. However, cpDNAs from all the five Populus species showed different restriction fragmentation pattern from that of tobacco with the four restriction enzymes tested. Southern hybridization with tobacco rbcL gene fragment as a probe also produced identical pattern among Populus species. The results indicate that cpDNAs in the genus are very well conserved during evolution.

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Triptolide Mimics the Effect of Dietary Restriction on Lifespan and Retards Age-related Diseases in Caenorhabditis elegans (트립톨라이드가 식이제한에 의한 수명연장과 노화관련 질환에 미치는 영향)

  • Beak, Sun-Mi;Park, Sang-Kyu
    • Journal of Life Science
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    • v.28 no.8
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    • pp.931-937
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    • 2018
  • Triptolide is a compound found in Tripterygium wilfordii and reported to have an anti-inflammatory and anti-oxidant activities. A previous study shows that the dietary supplementation with triptolide increases resistance to environmental stressors, including oxidative stress, heat shock, and ultraviolet irradiation, and extends lifespan in C. elegans. Here, we investigated the underlying mechanisms involved in the lifespan-extending effect of triptolide. The effect of triptolide on age-related diseases, such as diabetes mellitus and Alzheimer's disease, was also examined using animal disease models. The longevity phenotype conferred by triptolide was not observed in the eat-2 mutant, a well-known genetic model of dietary restriction, while there was an additional lifespan extension with triptolide in age-1 and clk-1 mutants. The long lifespan of age-1 mutant is resulted from a reduced insulin/IGF-1-like signaling and the clk-1 mutant lives longer than wild-type due to dysfunction of mitochondrial electron transport chain reaction. The effect of dietary restriction using bacterial dilution on lifespan also overlapped with that of triptolide. The toxicity of high glucose diet or transgenic human amyloid beta gene was significantly suppressed by the supplementation with triptolide. These findings suggest that triptolide can mimic the effect of dietary restriction on lifespan and onset of age-related diseases. We conclude that triptolide can be a strong candidate for the development of dietary restriction mimetics.

Regulation of GFP Expression Using the Tetracycline Inducible Retroviral Vector System (Tetracycline Inducible Retrovirus Vector System에 의한 GFP 유전자의 발현 조절)

  • Koo Bon Chul;Kwon Mo Sun;Kim Teoan
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.57-62
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    • 2005
  • One of the critical problems to be solved in transgenic animal production is uncontrollable constitutive expression of foreign genes, which usually results in serious physiological disturbances in the transgenic animal. To circumvent this problem, we constructed and tested two retrovirus vectors designed to express the GFP(green fluorescent protein) gene under the control of the tetracycline-inducible promoters. To maximize the GFP gene expression at turn-on state, WPRE(woodchuck hepatitis virus posttranscriptional regulatory element) sequence was introduced into the retrovirus vectors at downstream region of either the GFP gene or the sequence encoding rtTA(reverse tetracycline-controlled transactivator). Transformed cells were cultured in the medium supplemented with or without doxycycline(tetracycline derivative) for 48 hours, and induction efficiency was measured by comparing the GFP gene expression level using fluorometry and western blotting. Higher GFP expression was observed from the vector carrying the WPRE sequence at 3' side of the GFP gene, while tighter expression control(up to 20 fold) was obtained from the vector in which the WPRE sequence was placed at 3' side of rtTA sequence. The resulting tetracycline inducible vector system may be used in transgenic animal production and gene therapy.

Production of Supermale (YY) and Superfemale $({\Delta}YY)$ Nile Tilapia (Oreochromis niloticus) by Sex Reversal and Chromosome Manipulation I. Induction of Gynogenetic Diploid from XY Female (성전환 및 염색체 공학 기법을 이용한 초수컷(YY) 및 초암컷$({\Delta}YY)$ 나일틸라피아(Oreochromis niloticus) 생산 I. 성전환된 XY 암컷으로부터 자성발생성 이배체 유도)

  • Kim Dong Soo;Choi Yoon Hee;Noh Choong Hwan;Nam Yoon Kwon
    • Journal of Aquaculture
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    • v.8 no.4
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    • pp.295-306
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    • 1995
  • The present study was performed to produce supermales and superfemales in Oreochromis niloticus by combination of induced sex reversal and diploid gynogenesis. More than $95\%$ of female was obtained by oral administration of $17\beta-estradiol$ (480 mg/kg diet) to the eutheroembryonic larvae of this species. The result of progeny tests with XY pseudofemales showed that incidences of male progeny were ranged from $71.4\%\;to\;73.7\%$. p.opo.lion of male from XY pseudofemale was not significantly different (P>0.05) from that of $\chi^2$ (1:3), but it was significantly different (P<0.01) from that of $\chi^2$(1:1) Gynogenetic diploids were produced by applying cold shocks 3 min after insemination to the eggs of XY pseudofemale sperms were genetically inactivated by ultraviolet rays of 4,050 $erg/mm^2$. Fertilization rates of gynogenetic diploids were not different from that of their controls, however, hatching rates and early survival rates were silghtly lower than those of controls. In the cytogenetic studies, there was no difference in the size of cell and nucleus between the gynogenetic diploids and the controls. At 80 days after hatching, frequencies of gynogenetic male were higher than those of gynogenetic female (P<0.01) in their population.

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A Recombinant Microbial Biosensor for Cadmium and Lead Detection (카드뮴 및 납 검출을 위한 재조합 미생물 바이오센서)

  • Shin, Hae Ja
    • Journal of Life Science
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    • v.26 no.5
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    • pp.503-508
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    • 2016
  • Biosensors have been used as first-step monitoring tools to detect on-site samples in a simple and cost-effective manner. Numerous recombinant microbial biosensors have been exploited for monitoring on-site toxic chemicals and biological signals. Herein, a recombinant microbial biosensor was constructed for monitoring cadmium. The cadmium responding cadC regulatory gene and it’s promoter from Staphylococcus aureus was amplified through PCR, fused with the lacZ gene, and transformed into Escherichia coli BL21 (DE3) cells. In the presence of cadmium, the biosensor cells express β-galactosidase showing red color development with chlorophenol red β-galactopyranoside (CPRG) as the enzymatic substrate. The biosensor cells showed the best β-galactosidase activity after 3 hr induction with cadmium at pH 5 and a detection range from 0.01 μM to 10 mM cadmium with a linearity from 0.01 to 0.1 μM cadmium (y = 0.98 x + 0.142, R2 = 0.98). Among the heavy metals, cadmium and lead showed good responses, tin and cobalt showed medium responses, and mercury and copper showed no responses. The biosensor cells showed good responses to several waste waters similar to buffer solution, all spiked with cadmium. The biosensor described herein could be applied for on-site cadmium monitoring in a simple and cost-effective manner without sample pretreatments.

Dystrophin Degradation in Skeletal Muscles with Lipid Enrichment in Cattle (지방 침착률이 높은 식용소에서 나타난 골격근의 디스트로핀 소실)

  • Jeon, Sung-Hwan;Kim, Ah-Young;Lee, Eun-Mi;Lee, Eun-Joo;Hong, Il-Hwa;Hwang, Ok-Kyung;Jeong, Kyu-Shik
    • Journal of Life Science
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    • v.26 no.5
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    • pp.592-602
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    • 2016
  • This study investigated the muscular dystrophin levels in freely moving Australian cattle mainly fed grass, freely moving Korean cattle fed mainly a grain diet, and Korean cattle fed a grain diet but housed in a relatively limited space of a cow house. The total skeletal muscle specimens of 244 cattle were collected and immediately fixed in 10% neutral formalin. The same area was biopsied from the cattle in both countries. The findings showed that fatty infiltration is highly correlated with membrane-associated protein degradation in skeletal muscle, and that among several membrane-associated proteins, dystrophin showed the most significant reduction in expression in the cattle with fatty infiltration. Similarly, CD36 was more highly expressed in the cattle with fatty infiltration of skeletal muscle. Various breeding factors, such as oxidative stress; the presence of oxidized lipids in the diet; and environmental factors such as exercise, temperature and amount of time spent, may have critical effects on the degradation of normal cytoskeleton proteins, which are required for maintaining normal skeletal muscle architecture. Among the sarcolemma membrane-associated proteins, dystrophin is the most sensitive membrane protein that is involved muscular dystrophy and muscular degeneration. Thus, the present findings may be useful for studies on muscular dystrophy in humans or the pathogenesis of muscular diseases in animal models.