• Title/Summary/Keyword: Centromere

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Nucleotide Analysis of Phaffia rhodozyma DNA Fragment That Functions as ARS in Saccharomyces cerevisiae

  • Chung, Hee-Young;Hong, Min-Hee;Chun, Young-Hyun;Bai, Suk;Im, Suhn-Young;Lee, Hwanghee-Blaise;Park, Jong-Chun;Kim, Dong-Ho;Chun, Soon-Bai
    • Journal of Microbiology and Biotechnology
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    • v.8 no.6
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    • pp.650-655
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    • 1998
  • The chromosomal DNA fragment from Phaffia rhodozyma CBS 6938 which is able to autonomously replicate in the yeast Saccharomyces cerevisiae was cloned on an integrative URA3 plasmid. Its minimal fragment exhibiting autonomously replicating activiy in the S. cerevisiae gave a higher frequency transformation efficiency than that found for centromere-based plasmid, and enabled extrachromosoma1ly stable transmission of the plasmids in one copy per yeast cell under non-selective culture condition. The 836-bp DNA element lacked an ORF and did not contain any acceptable match to an ARS core consensus. Sequence analysis, however, displayed a cluster of three hairpin-Ioop-sequences with individual $\triangle {G_{25}}^{\circ}C$ free energy value of -10.0, -17.5, and -17.0 kcal. $mor^{-l}$as well as a 9-bp sequence with two base pair mismatches to the S. cerevisiae/E. coli gyrase-binding site. This 836-bp sequence also included one 7-bp sequence analogous to the core consensus of centromeric DNA element III (CDEIII) of S. cerevisiae, but CDEIII-like 7 bp sequence alone did not give a replicative function in this yeast.

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Genetical and Pathological Studies on the Mutant Mice as an Animal Model for Deafness Disease

  • Lee, Jeong-Woong;Lee, Eun-Ju;Lee, Hoon-Taek;Chung, Kil-Saeng;Ryoo, Zae-Young
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.48-48
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    • 2001
  • A new neurological mutant has been found in the ICR outbred strain mouse. Affected mice display profound deafness and a head-tossing and bidirectional circling behavior, showing an autosomal recessive mode of inheritance. It was, therefore, named cir/Kr with the gene symbol cir. The auditory tests identified clearly the hearing loss of the cir mice when compared to wild type mice. Pathological studies confirmed the developmental defects in the middle ear, cochlea, cochlear nerve, and semicircular canal areas, which were correlated to the abnormal behavior observed in the cir mice. Thus, cir mice may be useful as a model for studying inner ear abnormalities and deafness. We have constructed a genetic linkage map by positioning 14 microsatellite markers across the (cir) region and intraspecific backcross between cir and C57BL/6J mice. The cir mouse harbors an autosomal recessive mutation on mouse chromosome 9. The cir gene was mapped to a region between D9Mit116 and D9Mit38 Estimated distances between cir and D9Mit116, and between cir and D9Mit38 are 0.7 and 0.2 cM, respectively. The gene in order was defines : centromere-D9Mit182-D9Mit51/D9Mit79/D9Mit310-D9Mit212/D9Mit184-D9Mit116-cir-D9Mit38-D9Mit20-D9Mit243-D9Mit16-D9Mit55/D9Mit125-D9Mit281. The mouse map location of the cir locus appears to be in a region homologous to human 3q21. Our present date suggest that the nearest flanking marker D9Mit38 provides a useful anchor for the isolation of the cir gene in a yeast artificial chromosome contig.

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Increased Expression of a Chemically Synthesized Human Lysozyme Gene in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 합성된 Human Lysozyme 유전자의 발현증대)

  • 김기운;최선욱;이승철;백현동;황용일
    • Microbiology and Biotechnology Letters
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    • v.26 no.1
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    • pp.34-39
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    • 1998
  • We have already prepared a human lysozyme (HLY) structural gene from chemically synthesized 38 oligomers with high codon usage in Saccharomyces cerevisiae. For directing the synthesis and secretion of HLY in S. cerevisiae, two types of expression vectors, a YCp centromere-based vector, pHK101 and a YEp 2-$\mu\textrm{m}$ circle-based vector, pHK501 were constructed. With the resulting plasmids, we have confirmed that yeast transformant harboring pHK501 has more secreted HLY than pHK101-transformant by using a lysoplate and a turbidimetric assay. In flask cultivation, pHK501-transformant produced active HLY about 8 times (55 units/$m\ell$) higher than pHK101-transformant. From batch cultivation, the HLY productivity was obtained with 1.12 units/$m\ell$/h, corresponding to a 1.8-fold increase compared with flask fermentation. These results indicate that yeast transformant with pHK501 vector overexpressed and secreted HLY than that of YCp type vector.

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A Comparative Karyotype Study in Korean Squirrels. I Karyotype Analysis of Sciunis vulgaris coreae and Tamlas sibiricus asiaticus by Conventional Giemsa Staining and C-Banding Method (한국산 다람쥐 핵형의 비교연구 I.일반염색과 C-Banding방법에 의한 한국산 청서(Sciurus vulgaris corea) 와 다람쥐(Tamias sibiricus asiaticus)의 핵형 분석)

  • 김종봉;이희영
    • The Korean Journal of Zoology
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    • v.33 no.2
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    • pp.222-230
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    • 1990
  • The karyotypes of Korean Sciunis vulgaris coreas and Tamias sibiricus asiaticus were analyzed by conventional Giemsa staining and C-banding method. The diploid chromosome number (2n) of Sciunis vulgaris coreae 40 consisting of 6 metacentric, 8 submetacentric, 3 subtelocentric and 2 telocentric autosome pairs, submetacentric X and acrocentric or subtelocentric Y chromosome. The arm number (NF) of this species was obtained as 72, excluding the gonosomal arms. Tamias sibiricus asiaticus has a 2n of 38. The karyotype was represented by 3 metacentric, 4 submetacentric, 5 subtelocentric and 6 telocentric autosome paits and 2 sex chromosome. The X chromosome was submetacentric chromosome and the Y was the smallest chromosome with a median. The NF was 60. In S. vulgaris coreae constitutive heterochromatins were observed at the centromeres and telomeres. Constitutive heterochnomatins of T sibiricus asiaticus were primarily observed at the centromeres. These results suggested that non-Robensonian reanagenents and distribution of constitutive heterochromatin played an imporiant role in karyological differentiation of these species.

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Positional cloning in mice: a new mutant mouse, Sims (Sexual Immaturity, Megaencephaly, and Seizure)

  • Koo, S.K.;Jin, S.J.;Lee, K.S.;Oh, B.S.
    • Proceedings of the Zoological Society Korea Conference
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    • 1999.10b
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    • pp.31-31
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    • 1999
  • Characterization of mutant mice has been utilized as an animal model for the study of human inherited diseases. In addition to the pathogenesis stduy using the mutant mice, the mice have been used for the identification of the genes causing the phenotypes. Functional cloning and positional cloning are two approaches, depending on the phenotypes of the mutant mice. Though it takes a long time positional cloning has been well used to identify the gene of which function can not be presumed from the mouse phenotype. Recently by the advance of the molecular tools and the human genome project close to 10,000 genetic markers are developed to make the procedure faster. We obtained a new mutant mouse, sims, spontaneously arose and the affected mouse has a mild tremor and seizure was observed. Homozygote in either sex is sterile since uterus growth in female and seminal vesicle in male are not induced for the growth in puberty, implying the abnormal hormonal regulation during puberty. Supporting this, there is no detectable testosterone in the serum of the mutant male and the brain of the mutant is 30% heavier than littermate. To identify the location of the mutated gene, intraspecies cross to CAST/Ei was carried out and the 37 affected mice was analyzed for the linkage. The gene was mapped on chromosome 18, 20 cM from the centromere. More than 500 F2 progenies have been analyzed for the linkage and the locus becomes narrow within 3cM between Egrl and Fgf gene.f gene.

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Genetic diversity analysis in Chinese miniature pigs using swine leukocyte antigen complex microsatellites

  • Wu, Jinhua;Liu, Ronghui;Li, Hua;Yu, Hui;Yang, Yalan
    • Animal Bioscience
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    • v.34 no.11
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    • pp.1757-1765
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    • 2021
  • Objective: The swine leukocyte antigen (SLA) gene group, which is closely linked and highly polymorphic, has important biomedical significance in the protection and utilization of germplasm resources. However, genetic polymorphism analyses of SLA microsatellite markers in Chinese miniature pigs are limited. Methods: Eighteen pairs of microsatellite primers were used to amplify the SLA regions of seven miniature pig breeds and three wild boar breeds (n = 346) from different regions of China. The indexes of genetic polymorphism, including expected heterozygosity (He), polymorphic information content (PIC), and haplotype, were analyzed. The genetic differentiation coefficient (Fst) and neighbor-joining methods were used for cluster analysis of the breeds. Results: In miniature pigs, the SLA I region had the highest numbers of polymorphisms, followed by the SLA II and SLA III regions; the region near the centromere had the lowest number of polymorphisms. Among the seven miniature pig breeds, Diannan small-ear pigs had the highest genetic diversity (PIC value = 0.6396), whereas the genetic diversity of the Hebao pig was the lowest (PIC value = 0.4330). The Fst values in the Mingguang small-ear, Diannan small-ear, and Yunnan wild boars were less than 0.05. According to phylogenetic cluster analysis, the South-China-type miniature pigs clustered into one group, among which Mingguang small-ear pigs clustered with Diannan small-ear pigs. Haplotype analysis revealed that the SLA I, II, and III regions could be constructed into 13, 7, and 11 common haplotypes, respectively. Conclusion: This study validates the high genetic diversity of the Chinese miniature pig. Mingguang small-ear pigs have close kinship with Diannan small-ear pigs, implying that they may have similar genetic backgrounds and originate from the same population. This study also provides a foundation for genetic breeding, genetic resource protection, and classification of Chinese miniature pigs.

Observation of Mitotic Chromosome behavior according to Different Treatment Methods of DNA Methylation Inhibitor

  • Seong-Wook Kang;Ji-Yoon Han;Seong-Woo Cho
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.221-221
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    • 2022
  • Chromosome breakage occurred by DNA methylation inhibitor. Zebularine is known as DNA methylation inhibitor and suitable for water solubility among different DNA methylation inhibitors as 5-Azacytidine and 5-aza-2'-deoxycytidine. We used zebularine as mutagen according to different methods by roots absorption and seed imbibition. After zebularine treatment, DNA methylation inhibitor, we observed mitotic chromosome behavior what is different according to two different treatment methods. First, seed imbibition treatment in 1,000 μM of zebularine solution for 72 hours in dark conditions. The second treatment to seedlings of Keumkang was also treated in 1,000 μM of zebularine solution for 72 hours after germination. Root and shoot showed different elongations in each treatment. Root absorption treatment(3.01±0.48, 2.00±0.26) showed the shortest elongation in root and shoot than control(8.16±0.61, 4.03±0.48) and seed imbibition treatment(4.33±0.80, 2.48±0.36). It can be explained root tip meristematic cell activity was damaged by DNA methylation inhibitor. Primary root tips were collected in DW for 24 hours at low temperature(0℃) and fixed in fixation solution for 3 days to chromosome observation in mitosis. Mitotic index, chromosome structure and chromosome aberration were observed by phase-contrast microscope. Mitotic index of the control(0.29) showed twice mitotic cells as the treated groups(imbibition 0.15, absorption 0.14). Observation of chromosomes showed some short chromosomes and loosen chromosomes affected by zebularine. It is considered because of zebularine damage DNA in mitosis. We observed "gap by chromosome breakage" in chromosomes that have loose parts between centromere and telomere. It seems demethylation of zebularine occurs chromosome breakage.

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Chromosome Studies on Several Wild Sepcies of Drosophilidae (야생 초파리 수종의 염색체에 관한 연구)

  • 강영선;김영진;방규환
    • The Korean Journal of Zoology
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    • v.7 no.2
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    • pp.13-18
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    • 1964
  • The chromosomes of thirteen wild forms of Drosophila obtained from Kwangnung in Kyunggi Province, Korea were investigated with the ganglion cells of both male and female larvae using the aceto-lactic orcein squashed method. The male chromosome patterns of the species observed in the present study are summarized as follows:

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The Genetic Variation of Pinus densiflora and Pinus thunbergii by Giemsa C-banding (소나무 및 곰솔의 염색체(染色體) C-분염(分染)에 의한 유전변이(遺傳變異))

  • Park, Sang Jun;Son, Doo Sik
    • Journal of Korean Society of Forest Science
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    • v.80 no.4
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    • pp.383-392
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    • 1991
  • The genetic variation of Pinus densiflora and Pinus thunbergii by Giemsa C-banding was investigated and the results were as follows : 1. From Karyotype analysis of P. densiflora and P. thunbergii by Giemsa C-banding, somatic chromosome numbers of both species were 2n=24. 2. Chromosome of P. densiflora was M-type in arm ratio and they were no variation among individuals but variation in number and position of the secondary constriction and telomere banding among individuals. 3. P. thunbergii showed also M-type in arm ratio of chromosome, however, there was no variation in both number and position of the secondary constriction among individuals. 4. From chromosome C-banding, bands were appeared in the position of centromere and the secondary constriction in both P. densiflora and P. thunbergii. 5. In P. densiflora, the bands were shown on the secondary-constriction in chromosome No. 3, 4 and 7 of all individuals and the bands of the secondary constriction in chromosome No. 1, 2 and 5 showed variation among individuals. In chromosome No. 9, 10 and 11, the bands were shown in telomere and showed variation among individuals. 6. In P. thunbergii, the bands were shown on the secondary constriction in chromosome No. 2, 3, 7 and 8, and were shown no variation among individuals. There was no band on telomere. 7. The genetic variation by C-banding were shown in P. densiflora among individuals but no in P. thunbergii, and were shown on the secondary constriction in chromosome No. 4 of Pinus densiflora and in clnromosome No. 8 of Pinus thunbergii. These are the difference between the two species by C-banding.

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Induction of Two Types of Gynogenetic Diploid of Sweet Fish, Plecoglossus altivelis and Verification by Isozyme Marker (은어 2종류의 자성발생 2배체의 유도와 Isozyme 유전자에 의한 배수성의 확인)

  • 손진기
    • Development and Reproduction
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    • v.4 no.1
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    • pp.79-85
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    • 2000
  • This study was made to optimize the conditions needed to produce two types of gynogenetic diploids in the sweet fish, Plecoglossus altivelis. Firstly, ultraviolet (UV) ray doses between 3,000 erg to 14,000 erg/$\textrm{mm}^2$ were tested to inactivate sperm genetically. Based on the appearance of the haploid syndromes in the embryo, a dose of UV ray 6000~7000 erg was required to inactivate sperm genetically. Then, cold shock treatment at 1~2$^{\circ}C$ for 15~30 min were conducted to retain the 2nd polar body in inseminated egg. The best elapsed time before the start of the cold shock was examined between 5~8 min. The experiments in which began 5 min after insemination at 1~2$^{\circ}C$ during 17.5 min gave 21.2% survival rate and 89.7% normal eyed embryo rate. The gynogenetic diploid produced by suppression of the first cleavage, a considerably high number of heteroploids appeared and high mortality was observed at the metamorphosis stage, so further investigation is needed. The production of gynogenetic diploids were confirmed by GPI isozyme marker. The heterozygous type in Gpi-1 locus was observed in the meiotic-G2N as a result of gene-centromere recombination during meiosis. The heterozygous type was never observed in mitotic-G2N and showed segregation into two homozygous types at Gpi-1 locus.

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