• Title/Summary/Keyword: Artificial chromosome

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Construction of Chromosome-Specific BAC Libraries from the Filamentous Ascomycete Ashbya gossypii

  • Choi Sang-Dun
    • Genomics & Informatics
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    • v.4 no.2
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    • pp.80-86
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    • 2006
  • It is clear that the construction of large insert DNA libraries is important for map-based gene cloning, the assembly of physical maps, and simple screening for specific genomic sequences. The bacterial artificial chromosome (BAC) system is likely to be an important tool for map-based cloning of genes since BAC libraries can be constructed simply and analyzed more efficiently than yeast artificial chromosome (YAC) libraries. BACs have significantly expanded the size of fragments from eukaryotic genomes that can be cloned in Escherichia coli as plasmid molecules. To facilitate the isolation of molecular-biologically important genes in Ashbya gossypii, we constructed Ashbya chromosome-specific BAC libraries using pBeloBAC11 and pBACwich vectors with an average insert size of 100 kb, which is equivalent to 19.8X genomic coverage. pBACwich was developed to streamline map-based cloning by providing a tool to integrate large DNA fragments into specific sites in chromosomes. These chromosome-specific libraries have provided a useful tool for the further characterization of the Ashbya genome including positional cloning and genome sequencing.

Manipulation of Mini-Yeast Artificial Chromosome Containing Xylan Metabolism Related Genes and Mitotic Stability Analysis in Yeast (Xylan 대사유전자를가진미니효모인공염색체의가공및 Mitotic Stability 분석)

  • Da-In Kang;Yeon-Hee Kim
    • Microbiology and Biotechnology Letters
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    • v.50 no.3
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    • pp.436-440
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    • 2022
  • In this study, yeast artificial chromosome Insert (YAC) harboring genes which related xylan metabolism was constructed by using chromosome manipulation technique. For efficient chromosome manipulation, each splitting fragment (DNA module) required for splitting process was prepared and these DNA modules were transformed into Saccharomyces cerevisiae strain YKY164. By two-rounds chromosome splitting, yeast chromosome VII (1,124 kb) was split 887 kb-YAC, 45 kb-mini YAC and 198 kb-YAC and YKY183 strain containing 18 chromosomes was constructed. Splitting efficiency for chromosome manipulation was 50- 78% and expression level of foreign genes on 45 kb-mini YAC and enzyme activity were indistinguishable from that of the YKY164 strain. Furthermore, xylan-degraded products by recombinant enzymes were confirmed and mini-yeast artificial chromosome maintained stable mitotic stability without chromosome loss during 160 generations.

Simultaneous Overexpression of Integrated Genes by Copy Number Amplification of a Mini-Yeast Artificial Chromosome

  • Jung, Heo-Myung;Kim, Yeon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.821-825
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    • 2018
  • A copy number amplification system for yeast artificial chromosomes (YACs) was combined with simultaneous overexpression of genes integrated into a YAC. The chromosome VII (1,105 kb) was successfully split to 887 kb, 44 kb containing the element for copy number amplification, and a 184-kb split-YAC. The 44-kb split-mini YAC was amplified a maximum of 9-fold, and the activity of the reporter enzymes integrated into the split-mini YAC increased about 5-7-fold. These results demonstrate that the mini-YAC containing a targeted chromosome region can be readily amplified, and the specific genes in the mini-YAC could be overexpressed by increasing the copy number.

Chromosome Karyotype Classification using Multi-Step Multi-Layer Artificial Neural Network (다단계 다층 인공 신경회로망을 이용한 염색체 핵형 분류)

  • Chang, Yong-Hoon;Lee, Kwon-Soon;Chong, Hyeng-Hwan;Jun, Kye-Rok
    • Proceedings of the KOSOMBE Conference
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    • v.1995 no.11
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    • pp.197-200
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    • 1995
  • In this paper, we proposed the multi-step multi-layer artificial neural network(MMANN) to classify the chromosome, Which is used as a chromosome pattern classifier after learning. We extracted three chromosome morphological feature parameters such as centromeric index, relative length ratio, and relative area ratio by means of preprocessing method from ten chromosome images. The feature parameters of five chromosome images were used to learn neural network and the rest of them were used to classify the chromosome images. The experiment results show that the chromosome classification error is reduced much more, comparing with less feature parameters than that of the other researchers.

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The Implementation of Hierarchical Artificial Neural Network Classifier for Chromosome Karyotype Classification (염색체 핵형 분류를 위한 계층적 인공 신경회로망 분류기 구현)

  • Jeon, Gye-Rok;Choe, Uk-Hwan;Nam, Gi-Gon;Eom, Sang-Hui;Lee, Gwon-Sun;Jang, Yong-Hun
    • Journal of Biomedical Engineering Research
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    • v.18 no.3
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    • pp.233-241
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    • 1997
  • The research on chromosomes is very significant in cytogenetics since genes of the chromosomes control revelation of the inheritance plasma. The human chromosome analysis is widely used to study leukemia, malignancy, radiation hazard, and mutagen dosimetry as well as various congenital anomalies such as Down's, Klinefelter's, Edward's, and Patau's syndrome. The framing and analysis of the chromosome karyogram, which requires specific cytogenetic knowledge is most important in this field. Many researches on automated chromosome karyotype analysis methods have been carried out, some of which produced commercial systems. However, there still remains much room to improve the accuracy of chromosome classification and to reduce the processing time in real clinic environments. In this paper, we proposed a hierarchical artificial neural network(HANN) to classify the chromosome karyotype. We extracted three or four chromosome morphological feature parameters such as centromeric index, relative length ratio, relative area ratio, and chromosome length by preprocessing from ten human chromosome images. The feature parameters of five human chromosome images were used to learn HANN and the rest of them were used to classify the chromosome images. The experiment results show that the chromosome classification error is reduced much more than that of the other researchers using less feature parameters.

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Development of Simultaneous YAC Manipulation-Amplification (SYMA) system by Chromosome Splitting Technique Harboring Copy Number Amplification System (복제수 증폭시스템과 염색체 분단기술을 이용한 Simultaneous YAC Manipulation-Amplification (SYMA) 시스템의 개발)

  • Kim, Yeon-Hee;Nam, Soo-Wan
    • Journal of Life Science
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    • v.20 no.5
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    • pp.789-793
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    • 2010
  • Artificial chromosome manipulation and amplification of single-copy yeast artificial chromosome (YAC) are usually required in order to use YACs for applications such as physical mapping and functional analysis in eukaryotes. We designed and implemented a Simultaneous YAC Manipulation-Amplification (SYMA) system that combines the copy number amplification system of YAC with a convenient YAC manipulation system. To achieve the desired split and to amplify a YAC clone-harboring plant chromosome, a pBGTK plasmid containing a conditional centromere and thymidine kinase (TK) gene was constructed as a template to amplify the splitting fragment via PCR. By splitting, new 490-kb and 100-kb split YACs containing the elements for copy number amplification were simultaneously generated from a 590-kb YAC clone. The 100-kb split YAC was then successfully amplified 14.4-fold by adding 3 mg/ml sulfanilamide and $50\;{\mu}g/ml$ methotrexate (S3/M50) as inducing substances.

The Genetic Algorithm using Variable Chromosome with Chromosome Attachment for decision making model (의사결정 모델을 위한 염색체 비분리를 적용한 가변 염색체 유전 알고리즘)

  • Park, Kang-Moon;Shin, Suk-Hoon;Chi, Sung-Do
    • Journal of the Korea Society for Simulation
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    • v.26 no.4
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    • pp.1-9
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    • 2017
  • The Genetic Algorithm(GA) is a global search algorithm based on biological genetics. It is widely used in various fields such as industrial applications, artificial neural networks, web applications and defense industry. However, conventional Genetic Algorithm has difficulty maintaining feasibility in complicated situations due to its fixed number of chromosomes. This study proposes the Genetic Algorithm using variable chromosome with chromosome attachment. And in order to verify the implication of changing number of chromosomes in the simulation, it applies the Genetic Algorithm using variable chromosome with chromosome attachment to antisubmarine High Value Unit(HVU) escort mission simulation. As a result, the Genetic Algorithm using variable chromosome has produced complex strategies faster than the conventional method, indicating the increase of the number of chromosome during the process.

Influence of Chromosome Number on Cell Growth and Cell Aging in Yeast (효모에서 염색체의 수가 세포성장과 노화에 미치는 영향)

  • Kim, Yeon-Hee
    • Journal of Life Science
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    • v.26 no.6
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    • pp.646-650
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    • 2016
  • The influence of chromosome number on cell growth and cell aging was investigated in various yeast strains that have many artificial chromosomes constructed using a chromosome manipulation technique. Host strain FY833 and the YKY18, YKY18R, YKY24, and YKY30 strains harboring 16 natural chromosomes, 18 chromosomes, 18 chromosomes containing rDNA chromosome, 24 chromosomes, and 30 chromosomes, respectively, were used, and the specific growth rate of each strain was compared. The specific growth rates in the YKY18 and YKY24 strains were indistinguishable from that in the host strain, while those of the YKY18R and YKY30 strains were reduced to approximately 25% and 40% of the host strain level, respectively. Subsequently, the replicative life span was examined to investigate the relationship between the number of chromosomes and cell aging, and the life span was decreased to approximately 14% and 45% of the host strain level in the YKY24 and YKY30 strains, respectively. Moreover, telomere length, well known as a senescence factor, was shorter and more diversified in the strain, showing decreased life span. Therefore, these results suggest the possibility that an increase in the number of chromosomes containing artificial chromosomes caused cell aging, and we expected these observations would be applied to improve industrial strain harboring of versatile and special artificial chromosomes.

Creation of an Ethanol-Tolerant Yeast Strain by Genome Reconstruction Based on Chromosome Splitting Technology

  • Park, A-Hwang;Sugiyama, Minetaka;Harashima, Satoshi;Kim, Yeon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.184-189
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    • 2012
  • We sought to breed an industrially useful yeast strain, specifically an ethanol-tolerant yeast strain that would be optimal for ethanol production, using a novel breeding method, called genome reconstruction, based on chromosome splitting technology. To induce genome reconstruction, Saccharomyces cerevisiae strain SH6310, which contains 31 chromosomes including 12 artificial mini-chromosomes, was continuously cultivated in YPD medium containing 6% to 10% ethanol for 33 days. The 12 mini-chromosomes can be randomly or specifically lost because they do not contain any genes that are essential under high-level ethanol conditions. The strains selected by inducing genome reconstruction grew about ten times more than SH6310 in 8% ethanol. To determine the effect of mini-chromosome loss on the ethanol tolerance phenotype, PCR and Southern hybridization were performed to detect the remaining mini-chromosomes. These analyses revealed the loss of mini-chromosomes no. 11 and no. 12. Mini-chromosome no. 11 contains ten genes (YKL225W, PAU16, YKL223W, YKL222C, MCH2, FRE2, COS9, SRY1, JEN1, URA1) and no. 12 contains fifteen genes (YHL050C, YKL050W-A, YHL049C, YHL048C-A, COS8, YHLComega1, ARN2, YHL046W-A, PAU13, YHL045W, YHL044W, ECM34, YHL042W, YHL041W, ARN1). We assumed that the loss of these genes resulted in the ethanol-tolerant phenotype and expect that this genome reconstruction method will be a feasible new alternative for strain improvement.

The Origin of Artificial Species: Genetic Robot

  • Kim Jong-Hwan;Lee Kang-Hee;Kim Yong-Duk
    • International Journal of Control, Automation, and Systems
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    • v.3 no.4
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    • pp.564-570
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    • 2005
  • This paper provides a basis for investigating 'The Origin of Artificial Species,' as a robot can be considered as an artificial creature. To design an artificial creature, its general internal architecture is presented and its artificial chromosomes are proposed as its essential components. Rity as an artificial creature is developed in a virtual world of PC to test the world's first robotic 'chromosomes,' which are a set of computerized DNA (Deoxyribonucleic acid) codes for creating robots (artificial creatures) that can have their own personality, and can ultimately reproduce their kind, or even evolve as a distinct species. The effectiveness of the artificial chromosomes is demonstrated by implanting the genetic code into two Ritys living in a virtual world, in order to define their personality.