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Phylogenetic Analysis based on Metallothionein Gene Sequence of an Indigenous Species Pisidium (Neopisidium) coreanum in Korea (한국 고유종 Pisidium (Neopisidium) coreanum (산골조개) 의 metallothionein 유전자를 기초로 한 분자계통 분류학적 연구)

  • Baek, Moon-Ki;Lee, Jun-Seo;Kang, Se-Won;Lee, Jae-Bong;Kang, Hyun-Jung;Jo, Yong-Hun;Noh, Mi-Young;Han, Yeon-Soo;Choi, Sang-Haeng;Chae, Sung-Hwa;Park, Hong-Seog;Lee, Jun-Sang;Lee, Yong-Seok
    • The Korean Journal of Malacology
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    • v.25 no.2
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    • pp.153-160
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    • 2009
  • Pisidium (Neopisidium) coreanum is a freshwater snail and lives in spring water near mountain areas. Interestingly, this snail has been traditionally regarded as medicinal food, and thus has been used as folk remedies for healing broken bones. Recently, alpha classification on Pisidium (Neopisidium) coreanum through redescription has been conducted. However, not much attention has been made in beta classification. In this study, we performed the beta classification based on metallothionein (MT) genes found from various organisms. To this end, the complete cDNA sequences were obtained from the Expressed Sequence Tag (EST) sequencing project of Pisidium (Neopisidium) coreanum. The coding region (315 bp) encoded an amino acid sequence of 105 residues. The combined results from BLAST analyses, multiple sequence alignment and molecular phylogenetic study of Pc-MT gene indicate that Pisidium (Neopisidium) coreanum has similarity to freshwater bivalve such as Dreissena polymorpha (zebra mussel), Unio tumidus (swollen river mussel) and Crassostrea ariakensis (suminoe oyster).

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Effect of Trichostatin A on Anti HepG2 Liver Carcinoma Cells: Inhibition of HDAC Activity and Activation of Wnt/β-Catenin Signaling

  • Shi, Qing-Qiang;Zuo, Guo-Wei;Feng, Zi-Qiang;Zhao, Lv-Cui;Luo, Lian;You, Zhi-Mei;Li, Dang-Yang;Xia, Jing;Li, Jing;Chen, Di-Long
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7849-7855
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    • 2014
  • Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.

The First Report on the Acanthocephalan Infection of the Dybowskii's Brown Frogs (Rana dybowskii) Collected Inside and Outside the Commercial Frog Farms in Korea (국내 개구리 양식장 내·외에서 채집된 북방산개구리(Rana dybowskii)의 구두충 감염 최초보고)

  • Kim, Jong-Sun;Koo, Kyo-Soung;Park, Jae-Jin;Kwon, Sera;Choia, Woo-Jin;Cho, Han-Na;Park, Daesik
    • Korean Journal of Environment and Ecology
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    • v.30 no.4
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    • pp.694-704
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    • 2016
  • Various infectious and parasitic diseases are known to be the main factors that cause decline in the global amphibian population. In Korea, commercial frog farms have been running since 2005. However, until now, studies on diseases including studies on parasitic diseases that occur in farm frogs have not been conducted. In this study, we studied and compared the acanthocephalan infection rates and the number of parasites in the body cavity, stomach, and small and large intestines of the Dybowskii's brown frogs collected from inside and outside the frog farms in Inje, Goesan, Gongju and Boryeong. In addition, we classified the acanthocephalan parasites into genera by analyzing their nuclear 18S rRNA genes. On an average, 51.7% of the investigated frogs were infected by acanthocephalans, which belong to the Centrorhynchus genus. The infection rate of the frogs collected in the Inje farm was 15%, significantly lower than those from the Goesan, Gongju and Boryeong farms. The rate in Goesan was 55%, which is lower than Gongju (80%) and Boryeong (90%) although it is not statistically significant. No difference was found in the infection rate and in the number of parasites in male and female frogs and between the groups collected from inside and outside of the farms. The number of infected parasites negatively correlated with the body condition of the frogs. The most parasites were found in the stomach followed by the small and large intestines and the body cavity. This study is the first report on the Centrorhynchus acanthocephalan infection of amphibians in Korea and it suggests the necessity for acanthocephalan parasite management and for conducting further disease-related studies in commercial frog farms.

Effects of Cultivated Wild Panax ginseng Extract on the Proliferation, Differentiation and Mineralization of Osteoblastic MC3T3-E1 Cells (산양삼(cultivated wild Panax ginseng) 추출물이 조골세포 활성에 미치는 영향)

  • Seo, Hyun-Ju;Eo, Hyun Ji;Kim, Hyun Jun;Jeon, Kwon Seok;Park, Gwang Hun;Hong, Se Chul;Jeong, Jin Boo
    • Korean Journal of Plant Resources
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    • v.33 no.4
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    • pp.227-236
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    • 2020
  • Panax ginseng C.A. Meyer (P. ginseng) is known to exert a wide range of pharmacological effects both in vitro and in vivo. Although studies on ginsenoside, antioxidant activity, and anticancer effect of the cultivated wild Panax ginseng (CWP) have been conducted, there is little research on the effect of CWP extract on bone metabolism. In this study, we investigated the potential anti-osteoporotic properties of CWP on the growth and differentiation of MC3T3-E1 cells. CWP significantly increased the viability and proliferation of MC3T3-E1 cells. CWP activated intracellular alkaline phosphatase (ALP) activity in MC3T3-E1 cells. In addition, CWP increased the mineralized nodules in MC3T3-E1 cells. Furthermore, CWP increased the expression of genes such as Runx2, ALP, OPN and OCN associated with osteoblast growth and differentiation in a dose-dependent manner.

qVDT11, a major QTL related to stable tiller formation of rice under drought stress conditions

  • Kim, Tae-Heon;Cho, Soo-Min;Han, Sang-Ik;Cho, Jun-Hyun;Kim, Kyung-Min;Lee, Jong-Hee;Song, You-Chun;Park, Dong-Soo;Oh, Myung-Gyu;Shin, Dongjin
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.91-91
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    • 2017
  • Drought is the most serious abiotic stress limiting rice production. However, little progress has been made in the genetic analysis of drought tolerance, because it is a complex trait controlled by a number of genes and affected by various environmental factors. In here, we screened 218 rice genetic resources for drought tolerance at vegetative stage and selected 32 highly drought tolerant varieties in greenhouse. Under rain-fed conditions, Grain yield of Nagdong was decreased by 53.3% from 517 kg/10a to 241 kg/10a when compare to irrigation condition. By comparison, grain yield of Samgang was decreased by 23.6% from 550 kg/10a to 420 kg/10a. The variety Samgang exhibited strong drought tolerance and stable yield in rain-fed conditions and was selected for further study. To identify QTLs for drought tolerance, we examined visual drought tolerance (VDT) and relative water content (RWC) using a doubled haploid (DH) population consisted of 101 lines derived from a cross between Samgang (a drought tolerance variety) and Nagdong (a drought sensitive variety). Three QTLs for VDT were located on chromosomes 2, 6, and 11, respectively, and explained 41.8% of the total phenotypic variance. qVDT2, flanked by markers RM324 and S2016, explained 8.8% of the phenotypic variance with LOD score of 3.3 and an additive effect of -0.6. qVDT6 was flanked by S6022 and S6023 and explained 12.7% of the phenotypic variance with LOD score of 5.0 and an additive effect of -0.7. qVDT11, flanked by markers RM26765 and RM287, explained 19.9% of the phenotypic variance with LOD score of 7.1 and an additive effect of -1.0. qRWC11 was the only QTL for RWC to be identified; it was in the same locus as qVDT11. qRWC11 explained 19.6% of the phenotypic variance, with a LOD score of 4.0 and an additive effect of 9.7. To determine QTL effects on drought tolerance in rain-fed paddy conditions, seven DH lines were selected according to the number of QTLs they contained. Of the drought tolerance associated QTLs, qVDT2 and qVDT6 did not affect tiller formation, but qVDT11increased tiller number. Tiller formation was most stable when qVDT2 and qVDT11 were combined. DH lines with both of these drought tolerance associated QTLs exhibited the most stable tiller formation. These results suggest that qVDT11 is important for drought tolerance and stable tiller formation under drought stress condition in field.

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Novel quantitative trait loci for the strong-culm and high-yield related traits in rice detected from the F2 population between the super thick-culm and super grain-bearing line 'LTAT-29' and the high-yielding variety 'Takanari'

  • Nomura, Tomohiro;Yamamoto, Toshio;Ueda, Tadamasa;Yonemaru, Junichi;Abe, Akira;Adachi, Shunsuke;Hirasawa, Tadashi;Ookawa, Taiichiro
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.95-95
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    • 2017
  • Lodging is a serious issue in rice production, because it drastically decreases the biomass production and grain yield. Since the Green Revolution, the lodging resistance has been increased by lowering the moment of above-ground parts due to the short culm by the semi-dwarf gene sd1. However, it has been pointed out that sd1 alone has suppressive effects for biomass production and yield. To increase rice yield, the long-culm and large panicle type varieties with a superior lodging resistance need to be developed. To improve the lodging resistance and yield of these type varieties, it would be effective to identify novel alleles for these traits underlying natural variations in rice and to pyramid these alleles to a single rice variety. In order to perform this strategy, we have developed new rice lines derived from crosses among varieties with superior alleles. At first, TULT-gh-5-5 was selected from a cross between strong culm and high biomass variety Leaf Star and high-yielding variety Takanari, and TUAT-32HB was selected from a cross between high-yielding variety Akenohoshi and Takanari. Then, we developed the super thick-culm and super grain-bearing line, LTAT-29 derived from a cross between TULT-gh-5-5 and TUAT-32HB. In the current study, to identify the QTLs and genes relating to the strong culm and the high yield of LTAT-29, we performed QTL analysis using SNPs markers with $F_2$ population derived from a cross between LTAT-29 and Takanari. LTAT-29 has never lodged throughout the growth period despite it had long culms and heavy panicles. LTAT-29 had a larger outer diameter of the culm and twice the size of the section modulus than Takanari. As a result, the bending moment at breaking of LTAT-29 was significantly larger than that of Takanari. Brown rice yield of LTAT-29 was $9.2t\;ha^{-1}$ about 10% higher than that of Takanari due to the larger number of spikelets per panicle. LTAT-29 had a greater number of secondary branches per panicle. In the $F_2$ population between LTAT-29 and Takanari, we found continuous frequency distributions in the section modulus and the spikelet number per panicle. Two QTLs increased the section modulus by the alleles of LTAT-29 were detected on Chr.1L and Chr.2L. One QTL increased the spikelet number per panicle of Takanari by the allele of LTAT-29 was detected on Chr.1L, and two QTLs increased the number of secondary branches per panicle by the alleles of LTAT-29 were detected on Chr.1L and Chr.4L. It was found that the alleles of these QTLs were the japonica type originated from Leaf Star or Akenohoshi. The novel QTLs for the traits related to super thick-culm and super grain-bearing and their combinations could be utilized for improving the lodging resistance and yield in rice varieties.

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Construction of Tomato yellow leaf curl virus Clones for Resistance Assessment in Tomato Plants (토마토 작물의 TYLCV 저항성 평가에 이용할 수 있는 감염성 클론 개발)

  • Choi, Seung Kook;Choi, Hak Soon;Yang, Eun Young;Cho, In Sook;Cho, Jeom Deog;Chung, Bong Nam
    • Horticultural Science & Technology
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    • v.31 no.2
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    • pp.246-254
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    • 2013
  • Five isolates of Tomato yellow leaf curl virus (TYLCV) collected from various regions of Korea were amplified using PCR and determined the sequences of full-length genome, respectively. The PCR-amplified DNA of each TYLCV isolate was introduced into a binary vector to construct infectious clone containing 1.9 copies of the corresponding viral genome. Various cultivars and breeding lines of tomato were inoculated with Agrobacterium tumefaciens harboring infectious clone of each TYLCV isolate to assess resistance against TYLCV. Susceptible cultivar 'Super-sunread' revealed typical yellowing and narrowing of the upper leaves. In contrast, breeding linesTY12, GC9, GC171, and GC173, which contained the TY-1 and/or TY-3 genes that confer resistance against TYLCV in nature, were completely symptomless, suggesting that the lines were resistant to challenging TYLCV isolates. Symptoms of TYLCV in susceptible tomato cultivars are significantly different from those of TYLCV in the resistant tomato cultivars at 30 days after agroinfiltration. Although genomic DNAs of TYLCV were detected from the breeding lines TY12, GC9, GC171, and GC173 using real-time PCR analysis with specific primers, levels of TYLCV DNA accumulation in the resistant breeding lines were much lower than those of TYLCV DNA accumulation in susceptible tomato cultivars. Similar symptom severity and levels of TYLCV DNA accumulation were observed from TYLCV infections mediated by Bemisia tabaci in the resistant and susceptible tomato cultivars. Concentration of agrobacterium did not affect the response of tomato cultivars against TYLCV inoculation. Taken together, these results suggest that TYLCV inoculation via agroinfiltration is as effective as inoculation through Bemisia tabaci and is useful for breeding programs of TYLCV-resistant tomato.

Amended Soil with Biopolymer Positively Affects the Growth of Camelina sativa L. Under Drought Stress (가뭄 조건 하에서 바이오폴리머 혼합 토양이 Camelina sativa L.의 생장에 미치는 긍정적 영향)

  • Lim, Hyun-Gyu;Kim, Hyun-Sung;Lee, Hyeon-Sook;Sin, Jung-Ho;Kim, Eun-Suk;Woo, Hyo-Seop;Ahn, Sung-Ju
    • Ecology and Resilient Infrastructure
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    • v.5 no.3
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    • pp.163-173
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    • 2018
  • The biopolymer (BP) used in this study is mainly composed of xanthan gum and ${\beta}$-glucan derived from microorganism and has been introduced as a novel material for soil stabilization. However, the broad applicability of BP has been suggested in the field of geotechnical engineering while little information is available about the effects of BP on the vegetation. The goal of this study is to find the BP effects on the growth of Camelina sativa L. (Camelina) under drought condition. For more thorough evaluation of BP effects on the plant growth, we examined not only morphological but also physiological traits and gene expression patterns. After 25 days of drought treatment from germination in the soil amended with 0, 0.25, 0.5, and 1% BP, we observed that the BP concentration was strongly correlated the growth of Camelina. When plants were grown under drought stress, Camelina in 0.5% BP mixture showed better physiological parameters of the leaf stomatal conductance, electrolyte leakage and relative water content compared to those in control soil without BP. Plant recovery rate after re-watering was higher and the development of lateral root was lower in BP amended soil. RNA expression of Camelina leaf treated with/without drought for 7 and 10 days showed that aquaporin genes transporting solutes at bio-membrane, CsPIP1;4, 2;1, 2;6 and TIP1;2, 2;1, were induced more in the plants with BP amendment and drought treatment. These results suggest that the soil amended with BP has a positive effect on the transport of nutrients and waters into Camelina by improving water retention in soil under drought condition.

Analysis of Stress Response of Domestic Chicken Breeds for the Development of a New Synthetic Parent Stock (국산 종계 개발을 위한 토종 계통들의 스트레스 반응 정도 분석)

  • Sohn, Sea Hwan;Cho, Eun Jung;Park, Ji Ae;Hong, Young Ho;Kim, Chong Dae
    • Korean Journal of Poultry Science
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    • v.42 no.2
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    • pp.157-167
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    • 2015
  • We compared the degrees of stress response of 12 domestic purebred chicken strains that have been bred at National Institute of Animal Science, RDA, Korea since 1980. As a physiological marker of stress response, the expression levels of heat shock protein (HSP)-70, HSP-$90{\alpha}$, HSP-$90{\beta}$, hydroxyl-3-methyl-glutaryl coenzyme A reductase (HMGCR) genes and telomere length were measured by quantitative real-time polymerase chain reaction using the lymphocytes of 1,101 chickens. There was significant difference in HSP-70, HSP-$90{\alpha}$, HMGCR expression and telomere length among the strains. There was also significant difference in HSP-$90{\alpha}$, HSP-$90{\beta}$, and HMGCR expression between male and female chickens. Different age groups of chicken exhibited different expression levels of HSP-70, HSP-$90{\alpha}$ and telomere length. The results of the HSPs expression level suggested that, the strains of R, L and Y were highly resistant to stress, whereas the strains of S, O and W were susceptible to stress. Although the statistical differences in some of HSPs gene expression existed between genders, the HSP expression results varied in different strains that some opposed to the others, and there might be interaction between strains and genders, which conclude that there was no difference in stress response between male and female chickens. Moreover, despite of significant difference in some of HSPs expression level, it was considered that there was no difference in stress response between ages due to the inconsistent trends among HSP markers.

Mammalian Cloning by Nuclear transfer, Stem Cell, and Enzyme Telomerase (핵치환에 의한 cloning, stem cell, 그리고 효소 telomerase)

  • 한창열
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.423-428
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    • 2000
  • In 1997 when cloned sheep Dolly and soon after Polly were born, it had become head-line news because in the former the nucleus that gave rise to the lamb came from cells of six-year-old adult sheep and in the latter case a foreign gene was inserted into the donor nucleus to make the cloned sheep produce human protein, factor IX, in e milk. In the last few years, once the realm of science fiction, cloned mammals especially in livestock have become almost commonplace. What the press accounts often fail to convey, however, is that behind every success lie hundreds of failures. Many of the nuclear-transferred egg cells fail to undergo normal cell divisions. Even when an embryo does successfully implant in the womb, pregnancy often ends in miscarriage. A significant fraction of the animals that are born die shortly after birth and some of those that survived have serious developmental abnormalities. Efficiency remains at less than one % out of some hundred attempts to clone an animal. These facts show that something is fundamentally wrong and enormous hurdles must be overcome before cloning becomes practical. Cloning researchers now tent to put aside their effort to create live animals in order to probe the fundamental questions on cell biology including stem cells, the questions of whether the hereditary material in the nucleus of each cell remains intact throughout development, and how transferred nucleus is reprogrammed exactly like the zygotic nucleus. Stem cells are defined as those cells which can divide to produce a daughter cell like themselves (self-renewal) as well as a daughter cell that will give rise to specific differentiated cells (cell-differentiation). Multicellular organisms are formed from a single totipotent stem cell commonly called fertilized egg or zygote. As this cell and its progeny undergo cell divisions the potency of the stem cells in each tissue and organ become gradually restricted in the order of totipotent, pluripotent, and multipotent. The differentiation potential of multipotent stem cells in each tissue has been thought to be limited to cell lineages present in the organ from which they were derived. Recent studies, however, revealed that multipotent stem cells derived from adult tissues have much wider differentiation potential than was previously thought. These cells can differentiate into developmentally unrelated cell types, such as nerve stem cell into blood cells or muscle stem cell into brain cells. Neural stem cells isolated from the adult forebrain were recently shown to be capable of repopulating the hematopoietic system and produce blood cells in irradiated condition. In plants although the term$\boxDr$ stem cell$\boxUl$is not used, some cells in the second layer of tunica at the apical meristem of shoot, some nucellar cells surrounding the embryo sac, and initial cells of adventive buds are considered to be equivalent to the totipotent stem cells of mammals. The telomere ends of linear eukaryotic chromosomes cannot be replicated because the RNA primer at the end of a completed lagging strand cannot be replaced with DNA, causing 5' end gap. A chromosome would be shortened by the length of RNA primer with every cycle of DNA replication and cell division. Essential genes located near the ends of chromosomes would inevitably be deleted by end-shortening, thereby killing the descendants of the original cells. Telomeric DNA has an unusual sequence consisting of up to 1,000 or more tandem repeat of a simple sequence. For example, chromosome of mammal including human has the repeating telomeric sequence of TTAGGG and that of higher plant is TTTAGGG. This non-genic tandem repeat prevents the death of cell despite the continued shortening of chromosome length. In contrast with the somatic cells germ line cells have the mechanism to fill-up the 5' end gap of telomere, thus maintaining the original length of chromosome. Cem line cells exhibit active enzyme telomerase which functions to maintain the stable length of telomere. Some of the cloned animals are reported prematurely getting old. It has to be ascertained whether the multipotent stem cells in the tissues of adult mammals have the original telomeres or shortened telomeres.

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