• Title/Summary/Keyword: gene integration

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The Application of Machine Learning Algorithm In The Analysis of Tissue Microarray; for the Prediction of Clinical Status

  • Cho, Sung-Bum;Kim, Woo-Ho;Kim, Ju-Han
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.366-370
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    • 2005
  • Tissue microarry is one of the high throughput technologies in the post-genomic era. Using tissue microarray, the researchers are able to investigate large amount of gene expressions at the level of DNA, RNA, and protein The important aspect of tissue microarry is its ability to assess a lot of biomarkers which have been used in clinical practice. To manipulate the categorical data of tissue microarray, we applied Bayesian network classifier algorithm. We identified that Bayesian network classifier algorithm could analyze tissue microarray data and integrating prior knowledge about gastric cancer could achieve better performance result. The results showed that relevant integration of prior knowledge promote the prediction accuracy of survival status of the immunohistochemical tissue microarray data of 18 tumor suppressor genes. In conclusion, the application of Bayesian network classifier seemed appropriate for the analysis of the tissue microarray data with clinical information.

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Lily Pollen Growth in vitro and Agrobacterium-mediated GUS Gene Transformation via Vacuum-Infiltration

  • Park, In-Hae;Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.4 no.4
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    • pp.151-154
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    • 2002
  • Conditions for lily pollen growth in vitro and transformation were optimized. Active pollen tube development was achieved effectively in a medium containing 7% sucrose with pH adjusted to 5.7 at the temperature of 27$^{\circ}C$ for about 16-24 hours. Pollen growth was little impaired by the presence of kanamycin at concentration up to 100 mg/L. Pollen rains near the beginning of germination stage were more reliable for Agrobacterium-mediated GUS DNA transformation via vacuum infiltration lasted for 20-40 minutes. GUS DNA integration and its expression in fully developed pollen tubes could be confirmed by Southern blot hybridization, RT-PCR and histochemical staining.

Toxicoinformatics: The Master Key for Toxicogenomics

  • Lee, Wan-Sun;Kim, Yang-Seok
    • Molecular & Cellular Toxicology
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    • v.1 no.1
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    • pp.13-16
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    • 2005
  • The current vision of toxicogenomics is the development of methods or platforms to predict toxicity of un characterized chemicals by using '-omics' information in pre-clinical stage. Because each chemical has different ADME (absorption, distribution, mechanism, excretion) and experimental animals have lots of variation, precise prediction of chemical's toxicity based on '-omics' information and toxicity data of known chemicals is very difficult problem. So, the importance of bioinformatics is more emphasized on toxicogenomics than other functional genomics studies because these problems can not be solved only with experiments. Thus, toxicoinformatics covers all information-based analytical methods from gene expression (bioinformatics) to chemical structures (cheminformatics) and it also deals with the integration of wide range of experimental data for further extensive analyses. In this review, the overall strategy to toxicoinformatics is discussed.

Expression and Purification of Bacteriophage Lambda Integrase by Fusion Protein System (단백질 융합 시스템을 이용한 Bacteriophage Lambda Integrase의 발현 및 정제)

  • 이나영;유승구
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.784-788
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    • 1995
  • The lambda Integrase (Int) carries out site-specific recombination between the two partner DNA sequences, attachment P (attP) and attachment B (attB). In order to study the recombination mechanism, a large quantity of pure integrase is required. Then, we constructed an int gene inserted recombinant plasmid (pNYL3) by using the pQE31 HIS-Tag vector, and produced the fusion protein, 6xHIS-Int from the E. coli TG1 strain carrying the pNYL3 plasmid. The recombinant protein produced was purified by phosphocellulose and Ni$^{++}$-NTA affinity column chromatographies. The result of the in vitro recombination assay using the standard reaction mixture containing 6xHIS-Int and partially purified integration host factor (IHF) showed that the 6xHIS-Int tagged recombination Integrase had the full recombination activity.

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Utilization of Pollen Grain from Liluim cv. Casablanca as a Transient Gene Expression Host (유전자의 일시발현 분석용 숙주개발을 위한 카사블랑카백합(Lilium cv. Casablanca) 화분립의 이용)

  • Park, Hee-Sung
    • Applied Biological Chemistry
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    • v.47 no.4
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    • pp.430-433
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    • 2004
  • Lilium cv. Casablanca pollen grains stored at $-70^{\circ}C$ were grown in pollen germination medium with Agrobacterium tumefaciens LBA4404 cells harboring pBI121 for 18 hr at $27^{\circ}C$. Following this, cefotaxime (250 mg/L) was treated for 6 hr to eradicate the bacterial cells. Histochemical GUS analysis revealed that the transgenic pollen displayed deep blue color mostly from 12 hr after the co-cultivation. Presence of $200\;{\mu}M$ acetosyringone was determined not to be more effective for GUS transformation than its absence. GUS DNA integration in the transgenic pollen genomic DNA was clearly demonstrated by Southern blot analysis.

Construction of Expression Vector for Functional Analysis of Target Protein in Streptomyces sp.

  • Lee, Yong-Jik;Ryu, Jae-Ki;Kim, Hyun-Soo
    • Biomedical Science Letters
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    • v.18 no.1
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    • pp.29-34
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    • 2012
  • Streptomycetes are gram-positive filamentous bacteria that are well-known for producing a vast array of bioactive compounds, including more than 70 % of commercially important antibiotics. For the research about Streptomyces sp., the protoplast and electroporation transformation method have been the general techniques for the construction of transformants. However, these techniques have low efficiency and are time-consuming. Another option is intergenic conjugation, which is used for DNA transfer using methylation-deficient E. coli as a DNA donor to avoid the methylated-DNA-dependent restriction systems of actinomycetes. This conjugation method has been widely improved and applied to many other actinomycetes. In this research, an effective transformation procedure for the construction of expression vector by using gateway system was established to avoid limit of restriction enzyme site for cloning of target gene based on transconjugation by Escherichia coli ET12567/pUZ8002 with a pSET152 integration vector.

Design and Implementation of SOAP Servers Object Model for Gene Interaction Databases (유전자 상호작용 데이터베이스 SOAP서버 객체 모델의 설계 및 구현)

  • LEE HO IL;Yoo Seongjoon;Kim Minkyung
    • Journal of KIISE:Databases
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    • v.32 no.2
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    • pp.120-128
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    • 2005
  • Recently main Bioinformatics databases(DDBJ, ENSEMBL, KEGG, etc.) provide analysis tools and data using web services for the convenience of bioinformaticians. Thus, defining SOAP server objects and their methods are very important to provide services for web services. We define SOAP server objects for interaction databases such as BIND, MINT and DIP.

SNP Detection Using DNA Chip (DNA칩을 이용한 SNP의 검출)

  • Choi, Yong-Sung;Moon, Jong-Dae;Lee, Kyung-Sup
    • Proceedings of the KIEE Conference
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    • 2006.07c
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    • pp.1319-1321
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    • 2006
  • This research aims to develop the multiple channel electrochemical DNA chip that has the above characteristic and be able to solve the problems. At first, we fabricated a high integration type DNA chip array by lithography technology. It is able to detect a plural genes electrochemically after immobilization of a plural probe DNA and hybridization of non-labeling target DNA on the electrodes simultaneously. It suggested that this DNA chip could recognize the sequence specific genes. It suggested that multichannel electrochemical DNA microarray is useful to develop a portable device for clinical gene diagnostic system.

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Metagenomic analysis of viral genes integrated in whole genome sequencing data of Thai patients with Brugada syndrome

  • Suwalak Chitcharoen;Chureerat Phokaew;John Mauleekoonphairoj;Apichai Khongphatthanayothin;Boosamas Sutjaporn;Pharawee Wandee;Yong Poovorawan;Koonlawee Nademanee;Sunchai Payungporn
    • Genomics & Informatics
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    • v.20 no.4
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    • pp.44.1-44.13
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    • 2022
  • Brugada syndrome (BS) is an autosomal dominant inheritance cardiac arrhythmia disorder associated with sudden death in young adults. Thailand has the highest prevalence of BS worldwide, and over 60% of patients with BS still have unclear disease etiology. Here, we performed a new viral metagenome analysis pipeline called VIRIN and validated it with whole genome sequencing (WGS) data of HeLa cell lines and hepatocellular carcinoma. Then the VIRIN pipeline was applied to identify viral integration positions from unmapped WGS data of Thai males, including 100 BS patients (case) and 100 controls. Even though the sample preparation had no viral enrichment step, we can identify several virus genes from our analysis pipeline. The predominance of human endogenous retrovirus K (HERV-K) viruses was found in both cases and controls by blastn and blastx analysis. This study is the first report on the full-length HERV-K assembled genomes in the Thai population. Furthermore, the HERV-K integration breakpoint positions were validated and compared between the case and control datasets. Interestingly, Brugada cases contained HERV-K integration breakpoints at promoters five times more often than controls. Overall, the highlight of this study is the BS-specific HERV-K breakpoint positions that were found at the gene coding region "NBPF11" (n = 9), "NBPF12" (n = 8) and long non-coding RNA (lncRNA) "PCAT14" (n = 4) region. The genes and the lncRNA have been reported to be associated with congenital heart and arterial diseases. These findings provide another aspect of the BS etiology associated with viral genome integrations within the human genome.

Functional analysis of seaR protein identified from Saccharopolyspora erythraea (희소방선균의 seaR 단백질 발현을 통한 기능 분석)

  • Ryu, Jae Ki;Kwon, Pil-Seung;Lee, Hyeong Seon
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.39-47
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    • 2015
  • Secondary metabolism in actinomycetes has been known to be controlled by a small molecule, ${\gamma}$-butyrolactone autoregulator, the binding of which to each corresponding receptor leads to the regulation of the transcriptional expression of the secondary metabolites. We expected that expression of an autoregulator receptor or a pleiotropic regulator in a non-host was to be gained insight of effective production of new metabolic materials. In order to study the function of the receptor protein (seaR), which is isolated from Saccharopolyspora erythraea, we introduced the seaR gene to Streptomyces coelicolor A3(2) as host strains. An effective transformation procedure for S. coelicolor A3(2) was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a ${\varphi}C31$-derived integration vector, pSET152, which contained int, oriT, attP and $ermEp^*$ (erythromycin promotor). Therefore, the pEV615 was introduced into S. coelicolor A3(2) by conjugation and integrated at the attB locus in the chromosome of the recipients by the ${\varphi}C31$ integrase (int) function. Exconjugant of S. coelicolor A3(2) containing the seaR gene was confirmed by PCR and transcriptional expression of the seaR gene in the transformant was analyzed by RT-PCR. In case of S. coelicolor A3(2), a phenotype microarray was used to analyze the phenotype of transformant compared with wild type by seaR expression. After that, in order to confirm the accuracy of the results obtained from the phenotype microarray, an antimicrobial susceptibility test was carried out. This test indicated that sensitivity of the transformant was higher than wild type in tetracycline case. These results indicated that some biosynthesis genes or resistance genes for tetracycline biosynthesis in transformant might be repressed by seaR expression. Therefore, subsequent experiments, analysis of transcriptional pattern of genes for tetracycline production or resistance, are needed to confirm whether biosynthesis genes or resistance genes for tetracycline are repressed or not.