• Title/Summary/Keyword: Y chromosome

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Molecular Cloning and Bioinformatic Analysis of SPATA4 Gene

  • Liu, Shang-Feng;Ai, Chao;Ge, Zhong-Qi;Liu, Hai-Luo;Liu, Bo-Wen;He, Shan;Wang, Zhao
    • BMB Reports
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    • v.38 no.6
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    • pp.739-747
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    • 2005
  • Full-length cDNA sequences of four novel SPATA4 genes in chimpanzee, cow, chicken and ascidian were identified by bioinformatic analysis using mouse or human SPATA4 cDNA fragment as electronic probe. All these genes have 6 exons and have similar protein molecular weight and do not localize in sex chromosome. The mouse SPATA4 sequence is identified as significantly changed in cryptorchidism, which shares no significant homology with any known protein in swissprot databases except for the homologous genes in various vertebrates. Our searching results showed that all SPATA4 proteins have a putative conserved domain DUF1042. The percentages of putative SPATA4 protein sequence identity ranging from 30% to 99%. The high similarity was also found in 1 kb promoter regions of human, mouse and rat SPATA4 gene. The similarities of the sequences upstream of SPATA4 promoter also have a high proportion. The results of searching SymAtlas (http://symatlas.gnf.org/SymAtlas/) showed that human SPATA4 has a high expression in testis, especially in testis interstitial, leydig cell, seminiferous tubule and germ cell. Mouse SPATA4 was observed exclusively in adult mouse testis and almost no signal was detected in other tissues. The pI values of the protein are negative, ranging from 9.44 to 10.15. The subcellular location of the protein is usually in the nucleus. And the signal peptide possibilities for SPATA4 are always zero. Using the SNPs data in NCBI, we found 33 SNPs in human SPATA4 gene genomic DNA region, with the distribution of 29 SNPs in the introns. CpG island searching gives the data about CpG island, which shows that the regions of the CpG island have a high similarity with each other, though the length of the CpG island is different from each other.This research is a fundamental work in the fields of the bioinformational analysis, and also put forward a new way for the bioinformatic analysis of other genes.

Selenium Effect on the Frequency of SCEs Induced by Heavy Metals in Human Lymphocytes (Selenium이 mercury, cadmium 및 chromium에 의한 자매염색분체교환(姉妹染色分體交換)의 빈도(頻度)에 미치는 영향(影響))

  • Koh, Dai-Ha;Ki, No-Suk
    • Journal of Preventive Medicine and Public Health
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    • v.23 no.1 s.29
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    • pp.1-10
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    • 1990
  • The protective effect of sodium selenite($Na_2SeO_3$) against the cytogenetic toxicity of heavy metals was investigated on human whole-blood cultures in relation to induction of sister chromatid exchange (SCE) in secondary metaphase chromosome. Methylmercury chloride($CH_3HgCl$), cadmium chloride($CdCl_2$), potassium dichromate($K_2Cr_2O_7$), and sodium selenite caused to the typically dose-dependent increase in sister chromatid exchanges (SCEs) by the concentrations ranging from $0.3{\mu}M\;to\;10{mu}M$. However, the inductions of sister chromatid exchanges by methylmercury chloride or cadmium chloride were inhibited by the simultaneous addition of sodium selenite $1.2{mu}M$. The frequencies of SCE were decreased to the level of control in the molar ratios as 2:1, 1:1, 1:2, and 1:4 of selenium selenite vs. methylmercury chloride, and as 1:1 and 1:2 of selenium selenite vs. cadmium chloride, while the frequencies of SCE induced by potassium dichromate were not changed by the addition of sodium selenite in culture condition. Mitotic indices were decreased in the higher concentrations of chemicals and not significantly changed by the simultaneous addition of sodium selenite to the culture condition containing each chemicals.

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Agronomic Characteristics and Aromatic Compositions of Korean Wild Codonopsis lanceolata Collections Cultivated in Field (한국 야생더덕 수집종의 노지 재배시 생육 특성과 향기성분 조성)

  • 이승필;김상국;민기군;조지형;최부술;이상철;김길웅
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.2
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    • pp.188-199
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    • 1996
  • The native ecological environment and aromatic constituents of Korean wild Codonopsis lanceolata and one Japanese strain were investigated to find Codonopsis lanceolata strains showing high aromatics, and to know regional differences among these strains. The results were as follows : There were no remarkable differences among the Korean wild C. lanceolata strains in ecological environments. Recovery yield of essential oils was highest in Togyusan strain with 0.009%. Difference in protein band patterns among these strains was not recognized, and peroxidase and esterase pattern changes were appeared in different collected regions at the leaf and root tissues. Major aromatic constituents were 11 kinds of aliphatic alcohols such as trans-2-hexenal, 1-hexanol, cis-3-hexanol, and trans-2-hexanol. And Togyusan strain, Sobaeksan strain, and Kayasan strain have the highest aliphatic alcohols of plant essential oils. In particular, BHT(butylated hydroxytoluene), one of the antioxidants, was detected in Chirisan strains.

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The effect of chemicals and chilling treatments on the frequency of metaphase in root tips of some economic crops (화학약품과 저온처리가 작물근단세포의 Metaphase 출현빈도에 미치는 영향)

  • Mun-Hue Heu;Hye-Yong Kim
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.2
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    • pp.3-9
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    • 1964
  • Root tips of wheat, soybean, cotton and barley were treated with cold temperature (12 or 24 hrs in $0^{\circ}C$ or 2$^{\circ}C$), 8-Hyd.exyquinolin (2 or 10 hrs in 0.03 or 0.1%) and colcllicine (2 or 10 hrs in 0.2 or 1, 0%), and the frequency of metaphase were observed. The results were summarized as follow; 1. Chilling the seminal roots before or after sooting from the seed significantly increased tile number of mitotic cells and the rate of metaphase cells to mitotic cells. The optimal duration of chilling seemed to be differ depending on the kinds of plant and 24 hours to be too long except wheat so far examined here. 9. 8-Hydroxyquinolin treatment, about 2 hours in 0.03%, increased the rate of metaphase cells. The higher concentration and the longer treatment of this chemical caused the lower frequency of mitotic cells generally. 3. Colchicine treatment, 2 to 10 hrs in 0.2 to 1.0%, increased the frequency of mitotic cells and the rate of metaphase cells. Colchicine treatment was same or superior than any other treatments on the increase of metaphase cells. 4. All the treatments examined here caused chromosome contraction with most serious in colchicine following by 8-Hydroxyquinolin and chilling. 5. The feasibility of general application of chilling under the check of proper temperature and proper duration depending on the kinds of crops were discussed.

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Diversity Analysis of Japonica Rice using MITE-transposon Display (MITE-AFLP를 이용한 자포니카 벼의 다양성 검정)

  • Hong Seong-Mi;Kwon Soo-Jin;Oh Chang-Sik;Wessler Susan R.;Ahn Sang-Nag
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.3
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    • pp.259-268
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    • 2006
  • Miniature inverted transposable elements (MITEs) are abundant genomic components in plant including rice. MITE-transposon display (MITE-TD) is an Amplified Fragment Length Polymorphism (AFLP)-related technique based on MITE sequence. In this study, we used the MITE-AFLP for the analysis of diversity and relation-ship of the 114 japonica accessions. Of the several MITEs, the mPing family was applied to detect polymorphisms based on PCR amplification. The BfaI adaptor primer and the specific primer derived from mPing terminal inverted repeat (TIR) region were used to PCR amplification of 114 accessions. Nine primer pairs produced a total of 160 polymorphic bands. PIC values of the polymorphic bands generated by nine primer pairs ranged from 0.269 (BfaI + ACT) to 0.426 (BfaI + T). Each accession revealed a distinct fingerprint with two primer combinations, BfaI + G and BfaI + C. Cluster analysis using marker-based genetic similarity classified 114 accessions into five groups. MITE-AFLP markers were genetically mapped using a population of 80 BILs (BC1F7) derived from a cross between the rice accessions, Milyang 23 and Hapcheonaengmi 3. Eight of the markers produced with the primer pair BfaI + 0 were mapped on chromosomes 1, 2, 4, 5, 7, and 9. Considering that one MITE-AFLP marker on chromosome 7 was tightly linked to the Rc gene, the MITE-AFLP markers will be useful for gene tagging and molecular cloning.

Induction of Apomixis by Chemical Mutagen Treatment and Ovule Development in Inbreed lines of Corn (옥수수 자식계통들에서 화학적 돌연변이 유발성질 처리에 따른 apomixis 유발과 배주발생)

  • 이호진;최근진;김태훈
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.37 no.5
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    • pp.476-485
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    • 1992
  • M1 plants which were produced from seed soaking in chemical mutagen, EMS or NaN$_3$, appeared wide morphorogical variations such as dwarf, albino, twisted leaf, white streaked leaf, and purpled stem. In mutants of reproductive organs, there were monoecious plants such as female-flower plant and male-flower plant, multiple spikes, and steriled plants among M1 plants. Also, barren stalk was increased significantly in M1 plants. Ear bagging at ear initiation stage prevented seed set on cob in normal plants. In spite of ear bagging, M1 plants which had cobs with seed set was 3.9-11.2% of stalks developed from seeds soaking with mutagens, but only three or four kernels could be matured on a cob. Ear bagging after mutagen injection into initiating ear produced 5.1-10% in cobs with seed set, but only 1.7-6.3 kernels could be matured. Cobs removed silk at four hours after artificial pollination increased the rate of cobs with seed set to 27%. Microscopic observation confirmed that ontogeny of kernels matured from ear bagging and mutagen treatment would be both adventitious and diplosporous apomictic reproduction. Chromosome set of M2 seedling was found to be diploid type in chromosomal counting of root tip. As M$_2$ plants showed an uniform appearence within each lines and their CV of plant height were ranged 4-6% in each lines, we concluded that they were apomictic progeny. But we could not find any marker traits combined with apomixis.

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Detection of Mitotic Centromere-Associated Kinesin (MCAK) During Cell-Cycle Progression of Human Jurkat T Cells Using Polyclonal Antibody Raised Against Its N- Terminal Region Overexpressed in E. coli

  • Jun, Do-Youn;Rue, Seok-Woo;Kim, Byung-Woo;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.912-918
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    • 2003
  • Mitotic centromere-associated kinesin (MCAK), which is a novel kinesin with a central motor domain, is believed to playa role in mitotic segregation of chromosome during the M phase of the cell cycle. In the present study, it is shown that a rabbit polyclonal antibody has been produced using the N-terminal region (187 aa) of human MCAK expressed in E. coli as the antigen. To express the N-terminal region in E. coli, the MCAK cDNA fragment encoding N-terminal 187 aa was obtained by PCR and was then inserted into the pET 3d expression vector. Molecular mass of the N-terminal region overexpressed in the presence of IPTG was 23.2 kDa on SDS-PAGE, and the protein was insoluble and mainly localized in the inclusion body that could be easily purified from the other cellular proteins. The N-terminal region was purified by electro-elution from the gel after the inclusion body was resolved on the SDS-PAGE. The antiserum obtained after tertiary immunization with the purified protein specifically recognized HsMCAK when subjected to Western blot analysis, and showed a fluctuation of the protein level during the cell cycle of human Jurkat T cells. Synchronization of the cell-cycle progression required for recovery of cells at a specific stage of the cell cycle was performed by either hydroxyurea or nocadazole, and subsequent release from each blocking at 2, 4, and 7 h. Northern and Western analyses revealed that both mRNA and protein of HsMCAK reached a maximum level in the S phase and declined to a basal level in the G1 phase. These results indicate that a polyclonal antibody raised against the N-terminal region (187 aa) of HsMCAK, overexpressed in E. coli, specifically detects HsMCAK (81 kDa), and it can analyze the differential expression of HsMCAK protein during the cell cycle.

Comparison of Methods for Stable Simultaneous Expression of Various Heterologous Genes in Saccharomyces cerevisiae (출아효모에서 다양한 이종 유전자의 안정적 동시발현을 위한 방법의 비교)

  • Jung, Heo-Myung;Kim, Yeon-Hee
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.667-672
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    • 2019
  • We compared two integration systems for stable expression of heterologous genes in Saccharomyces cerevisiae. A Candida glabrata-derived gene was used as the selective marker for the Cre/loxP system, and XYLP, XYLB, GRE3, and XYL2 genes were used as model heterologous genes and ligated into the universal pRS-CMT vector. The resulting pRS-XylP, pRS-XylB, pRS-Gre3, and pRS-Xyl2 plasmids were sequentially integrated into yeast chromosome VII by four integration processes (marker rescue and gene integration). The four introduced genes were successfully expressed. Further, the pRS-PBG2 plasmid harboring expression cassettes for the four genes was constructed for one-step integration. The four genes that were introduced were stably maintained as a gene cluster and were simultaneously expressed. The one-step integration was more effective for the simultaneous integration and expression of the four genes related to xylan/xylose metabolism. This method will enable the generation of a useful biosystem through appropriate use of gene integration methods.

Mitotic-Specific Methylation in the HeLa Cell through Loss of DNMTs and DMAP1 from Chromatin

  • Kim, Kee-Pyo;Kim, Gun-Do;Kang, Yong-Kook;Lee, Dong-Seok;Koo, Deog-Bon;Lee, Hoon-Taek;Chung, Kil-Saeng;Lee, Kyung-Kwang;Han, Yong-Mahn
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.27-27
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    • 2003
  • A diversified and concentrative approach of methylation player can be one of the most powerful studies in the understanding of global epigenetic modifications. Previous studies have suggested that DNA methylation contributes to transcriptional silencing through the several DNA methylation-mediated repression systems by hypermethylation, including methyltransferases (DNMTs), DNA methyltransferase association protein 1 (DMAPl), methyl-CpG binding domain (MBD), and histone deacetylases (HDACs). Assembly of these regulatory protein complexes act sequentially, reciprocally, and interdependently on the newly composed DNA strand through S phase. Therefore, these protein complexes have a role in coupling DNA replication to the designed turn-off system in genome. In this study, we attempted to address the role of DNA methylation by the functional analysis of the methyltransferase molecule, we described the involvement of DMAP1 and DNMTs in cell divistion and the effect of their loss. We also described distinct patterns that DMAP1 and DNMTs are spatially reorganized and displaced from condensing chromosomes as cells progress through mitosis in HeLa cell, COS7, and HIH3T3 cell cycle progressions. DNMT1, DNMT3b, and DMAP1 do not stably contact the genetic material during chromosome compaction and repressive expression. These finding show that the loss of activities of DNMTs and DMAP1 occure stage specifically during the cell cycle, may contribute to the integral balance of global DNA methylation. This is consistent with previous studies resulted in decreased histone acetyltransferases and HDACs, and differs from studies resulted in increased histone methyltransferases. Our results suggest that DNA methylation by DNMTs and DMAP1 during mitosis acts to antagonize hypermethylation by which this mark is epigenetical mitotic-specific methylation.

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Implementation of Evolving Neural Network Controller for Inverted Pendulum System (도립진자 시스템을 위한 진화형 신경회로망 제어기의 실현)

  • 심영진;김태우;최우진;이준탁
    • Journal of the Korean Institute of Illuminating and Electrical Installation Engineers
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    • v.14 no.3
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    • pp.68-76
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    • 2000
  • The stabilization control of Inverted Pendulum(IP) system is difficult because of its nonlinearity and structural unstability. Futhermore, a series of conventional techniques such as the pole placement and the optimal control based on the local linearizations have narrow stabilizable regions. At the same time, the fine tunings of their gain parameters are also troublesome. Thus, in this paper, an Evolving Neural Network Controller(ENNC) which its structure and its connection weights are optimized simultaneously by Real Variable Elitist Genetic Algorithm(RVEGA) was presented for stabilization of an IP system with nonlinearity. This proposed ENNC was described by a simple genetic chromosome. And the deletion of neuron, the according to the various flag types. Therefore, the connection weights, its structure and the neuron types in the given ENNC can be optimized by the proposed evolution strategy. And the proposed ENNC was implemented successfully on the ADA-2310 data acquisition board and the 80586 microprocessor in order to stabilize the IP system. Through the simulation and experimental results, we showed that the finally acquired optimal ENNC was very useful in the stabilization control of IP system.

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