• Title/Summary/Keyword: Ltd gene

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Multiplex TaqMan qPCR Assay for Detection, Identification, and Quantification of Three Sclerotinia Species

  • Dong Jae Lee;Jin A Lee;Dae-Han Chae;Hwi-Seo Jang;Young-Joon Choi;Dalsoo Kim
    • Mycobiology
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    • v.50 no.5
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    • pp.382-388
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    • 2022
  • White mold (or Sclerotinia stem rot), caused by Sclerotinia species, is a major air, soil, or seed-transmitted disease affecting numerous crops and wild plants. Microscopic or culture-based methods currently available for their detection and identification are time-consuming, laborious, and often erroneous. Therefore, we developed a multiplex quantitative PCR (qPCR) assay for the discrimination, detection, and quantification of DNA collected from each of the three economically relevant Sclerotinia species, namely, S. sclerotiorum, S. minor, and S. nivalis. TaqMan primer/probe combinations specific for each Sclerotinia species were designed based on the gene sequences encoding aspartyl protease. High specificity and sensitivity of each probe were confirmed for sclerotium and soil samples, as well as pure cultures, using simplex and multiplex qPCRs. This multiplex assay could be helpful in detecting and quantifying specific species of Sclerotinia, and therefore, may be valuable for disease diagnosis, forecasting, and management.

Inducible Expression of the Lactadherin Gene with a Reverse Tetracycline-Regulated Retroviral Vector System (Tetracycline으로 발현이 유도되는 Retrovirus Vector System을 이용한 Human Lactadherin 유전자의 전이와 발현)

  • 이용석;오훈규;권모선;박창식;김태완;박재복
    • Korean Journal of Animal Reproduction
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    • v.27 no.3
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    • pp.259-268
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    • 2003
  • Lactadherin (formerly known as BA46), a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and breast carcinoma cells and is known to prevent symptomatic rotavirus infections. In this study, we tried to transfer the human lactadherin gene to the Chinese Hamster Ovary (CHO) cells using retrovirus vector system and tested inducible expression of the gene under the tetracycline-controllable promoter. At first, tetracycline-mediated inducibility was tested using E.coli LacZ marker gene. NIH3T3 cells co-infected with RevTet-On and RevTRE-LacZ retrovirus vectors showed that the cells responded to doxycycline (a derivative of tetracycline) in a dose-dependent manner, and prominent induction of the lacZ gene expression was observed from 1 $\mu\textrm{g}$/ml of doxycycline concentration. Based on the results of the pilot experiment, inductional expression of the human lactadherin gene was conducted using RevTet-On and RevTRE-Ltd retrovirus vectors. Analysis with the RT-PCR demonstrated successful inductional expression of the lactadherin gene in the Chinese Hamster Ovary (CHO) cells. Considering that constitutive overexpression of the exogenous genes in the target cells causes serious physiological imbalance, the results obtained in this study will be very useful especially in the studies of gene therapy and transgenic animal production.

Development of Rapid Antibody-based Therapeutic Platform Correspondence for New Viruses Using Antigen-specific Single Cell Memory B Cell Sorting Technology (항원 특이적 단일 기억 B 세포 분리를 이용한 신종 바이러스 대응 신속 항체 플랫폼 개발)

  • Jiyoon Seok;Suhan Jung;Ye Gi Han;Arum Park;Jung Eun Kim;Young Jo Song;Chi Ho Yu;Hyeongseok Yun;Se Hun Gu;Seung-Ho Lee;Yong Han Lee;Gyeunghaeng Hur;Woong Choi
    • Journal of the Korea Institute of Military Science and Technology
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    • v.27 no.1
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    • pp.116-125
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    • 2024
  • The COVID-19 pandemic is not over despite the emergency use authorization as can see recent COVID-19 daily confirmed cases. The viruses are not only difficult to diagnose and treat due to random mutations, but also pose threat human being because they have the potential to be exploited as biochemical weapons by genetic manipulation. Therefore, it is inevitable to the rapid antibody-based therapeutic platform to quickly respond to future pandemics by new/re-emerging viruses. Although numerous researches have been conducted for the fast development of antibody-based therapeutics, it is sometimes hard to respond rapidly to new viruses because of complicated expression or purification processes for antibody production. In this study, a novel rapid antibody-based therapeutic platform using single B cell sorting method and mRNA-antibody. High immunogenicity was caused to produce antibodies in vivo through mRNA-antigen inoculation. Subsequently, antigen-specific antibody candidates were selected and obtained using isolation of B cells containing antibody at the single cell level. Using the antibody-based therapeutic platform system in this study, it was confirmed that novel antigen-specific antibodies could be obtained in about 40 days, and suggested that the possibility of rapid response to new variant viruses.

An information-theoretical analysis of gene nucleotide sequence structuredness for a selection of aging and cancer-related genes

  • Blokh, David;Gitarts, Joseph;Stambler, Ilia
    • Genomics & Informatics
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    • v.18 no.4
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    • pp.41.1-41.8
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    • 2020
  • We provide an algorithm for the construction and analysis of autocorrelation (information) functions of gene nucleotide sequences. As a measure of correlation between discrete random variables, we use normalized mutual information. The information functions are indicative of the degree of structuredness of gene sequences. We construct the information functions for selected gene sequences. We find a significant difference between information functions of genes of different types. We hypothesize that the features of information functions of gene nucleotide sequences are related to phenotypes of these genes.

Comparative Analysis of Completely Sequenced Insect Mitochondrial Genomes

  • Lee, Jin-Sung;Kim, Ki-Hwan;Suh, Dong-Sang;Park, Jae-Heung;Suh, Ji-Yoeun;Chung, Kyu-Hoi;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.1
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    • pp.1-6
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    • 2001
  • This paper reports a few characteristics of seven insect mitochondrial genomes sequenced completely (Bombyx mori, Drosophila melanogaster, D. yakuba, Apis mellifera, Anopheles gambiae, A. quadrimaculatus, and Locusta migratoria). Comparative analysis of complete mt genome sequences from several species revealed a number of interesting features (base composition, gene content, A+T-rich region, and gene arrangement, etc) of insect mitochondrial genome. The properties revealed by our work shed new light on the organization and evolution of the insect mitochondrial genome and more importantly open up the way to clearly aimed experimental studies for understanding critical roles of the regulatory mechanisms (transcription and translation) in mitochondrial gene expression.

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Inductional Expression of the Human Lactadherin Gene in Mouse Mammary Epithelial Cells

  • Kwon, Mo-Sun;Koo, Bon-Chul;Kim, Teoan
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.94-94
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    • 2002
  • Lactadherin (formerly known as BA46), a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and breast carcinomas and may prevent symptomatic rotavirus infections. In this study, under the control of mouse whey acidic protein (WAP) promoter, the expression pattern of lactadherin (Ltd) in lactogenic hormone-dependent mouse mammary epithelial cell line HC11 were tested. pLNWLtd construct containing 2.4 kilobases of the WAP promoter and 1.5 kilobases of human lactadherin gene was stably transfered into HC11 cells using retroviral vector system. Integration and expression level of the transgene was estimated using PCR and RT-PCR, respectively. Prominent induction of Ltd gene under the WAS promoter was accomplished in the presence of insulin, hydrocortisone and prolactin, while induction with insulin alone resulted in lower expression. Our results demonstrate that the expression of the transgene is increased by synergistic effect of several lactogenic hormones, including insulin, hydrocortisone, and prolactin.

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