• Title/Summary/Keyword: Target DNA

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Sequence-specific interaction between ABD-B homeodomain and castor gene in Drosophila

  • Kim, Keon-Hee;Yoo, Siuk
    • BMB Reports
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    • v.47 no.2
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    • pp.92-97
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    • 2014
  • We have examined the effect of bithorax complex genes on the expression of castor gene. During the embryonic stages 12-15, both Ultrabithorax and abdominal-A regulated the castor gene expression negatively, whereas Abdominal-B showed a positive correlation with the castor gene expression according to real-time PCR. To investigate whether ABD-B protein directly interacts with the castor gene, electrophoretic mobility shift assays were performed using the recombinant ABD-B homeodomain and oligonucleotides, which are located within the region 10 kb upstream of the castor gene. The results show that ABD-B protein directly binds to the castor gene specifically. ABD-B binds more strongly to oligonucleotides containing two 5'-TTAT-3' canonical core motifs than the probe containing the 5'-TTAC-3' motif. In addition, the sequences flanking the core motif are also involved in the protein-DNA interaction. The results demonstrate the importance of HD for direct binding to target sequences to regulate the expression level of the target genes.

Trend and Technology of Gene and Genome Research (유전자 및 유전체 연구 기술과 동향)

  • 이진성;김기환;서동상;강석우;황재삼
    • Journal of Sericultural and Entomological Science
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    • v.42 no.2
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    • pp.126-141
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    • 2000
  • A major step towards understanding of the genetic basis of an organism is the complete sequence determination of all genes in target genome. The nucleotide sequence encoded in the genome contains the information that specifies the amino acid sequence of every protein and functional RNA molecule. In principle, it will be possible to identify every protein resposible for the structure and function of the body of the target organism. The pattern of expression in different cell types will specify where and when each protein is used. The amino acid sequence of the proteins encoded by each gene will be derived from the conceptional translation of the nucleotide sequence. Comparison of these sequences with those of known proteins, whose sequences are sorted in database, will suggest an approximate function for many proteins. This mini review describes the development of new sequencing methods and the optimization of sequencing strategies for whole genome, various cDNA and genomic analysis.

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Trends in the development of discriminating between Angelica L. species using advanced DNA barcoding techniques (진보된 DNA barcoding 기술을 이용한 당귀(Angelica)속 식물의 기원 판별 기술에 관한 연구 동향)

  • Lee, Shin-Woo;Shin, Yong-Wook;Kim, Yun-Hee
    • Journal of Plant Biotechnology
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    • v.48 no.3
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    • pp.131-138
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    • 2021
  • We reviewed current research trends for discriminating between species of the Angelica genus, a group of important medicinal plants registered in South Korea, China, and Japan. Since the registered species for medicinal purposes differ by country, they are often adulterated as well as mixed in commercial markets. Several DNA technologies have been applied to distinguish between species. However, one of the restrictions is insufficient single-nucleotide polymorphisms (SNPs) within the target DNA fragments; in particular, among closely-related species. Recently, amplification refractory mutation system (ARMS)-PCR and highresolution melting (HRM) curve analysis techniques have been developed to solve such a problem. We applied both technologies, and found they were able to discriminate several lines of Angelica genus, including A. gigas Nakai, A. gigas Jiri, A. sinensis, A. acutiloba Kitag, and Levisticum officinale. Furthermore, although the ITS region differs only by one SNP between A. gigas Nakai and A. gigas Jiri, both HRM and ARMS-PCR techniques were powerful enough to discriminate between them. Since both A. gigas Nakai and A. gigas Jiri are native species to South Korea and are very closely related, they are difficult to discriminate by their morphological characteristics. For practical applications of these technologies, further research is necessary with various materials, such as dried or processed materials (jam, jelly, juice, medicinal decoctions, etc.) in commercial markets.

Distribution of the extended-spectrum beta-lactamase genes derived from microorganisms in the waterfront environments (주변 수계에서 미생물유래 extended-spectrum beta-lactamase 유전자의 분포)

  • Young-Min Bae
    • Journal of the Korean Applied Science and Technology
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    • v.39 no.6
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    • pp.916-923
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    • 2022
  • Water samples were collected from three spots(Namcheon, Changwoncheon and Cheongwoonji) in Changwon and genomic DNA was isolated from them. Quantitative PCR was performed with the isolated DNA as template and primers targeting five different class A extended-spectrum beta-lactamase(ESBL) genes(blaOXA-1, blaSHV, blaTEM, blaCTX-M-1, blaCTX-M-9). The number of total ESBL genes from each sample showed large variations between each sample. Thirty nanograms of DNA from Namcheon contained 1.93×106 copies of ESBL genes whereas the same amount of DNA from Changwoncheon contained 1.47×105 copies of ESBL genes. However, the same amount of DNA from Cheongwoonji pond contained only 9.5×103 copies of ESBL genes. The ratio of each ESBL genes showed little difference between Namcheon river and Changwoncheon river, but DNA from Cheongwoonji pond showed a large difference from the rest. blaOXA-1 gene was present at 65.3%, and blaCTX-M-1 gene 33.6% for Namcheon comprising together almost 99%. blaOXA-1 gene was present at 64.1%, and blaCTX-M-1 gene 19.1% for Changwoncheon comprising together over 83%. blaCTX-M-1 gene was present at 87.5% and blaCTX-M-9 genes 9.8% for Cheongwoonji, a pond which is a closed and isolated environment.

Simultaneous Detection of Listeria monocytogenes, Escherichia coli O157:H7, Bacillus cereus, Salmonella spp., and Staphylococcus aureus in Low-fatted Milk by Multiplex PCR

  • Kim, Ji-Hyun;Rhim, Seong-Ryul;Kim, Kee-Tae;Paik, Hyun-Dong;Lee, Joo-Yeon
    • Food Science of Animal Resources
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    • v.34 no.5
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    • pp.717-723
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    • 2014
  • A rapid and specific PCR assay for the simultaneous detection of Listeria monocytogenes, Escherichia coli O157:H7, Bacillus cereus, Salmonella spp., and Staphylococcus aureus in foods was developed to reduce the detection time and to increase sensitivity. Multiplex PCR developed in this study produced only actA, fliC, hbl, invA, ileS amplicons, but did not produce any non-specific amplicon. The primer sets successfully amplified the target genes in the multiplex PCR without any non-specific or additional bands on the other strains. The multiplex PCR assays also amplified some target genes from five pathogens, and multiplex amplification was obtained from as little as 1 pg of DNA. According to the results from the sensitivity evaluation, the multiplex PCR developed in this study detected 10 cells/mL of the pathogens inoculated in milk samples, respectively. The results suggested that multiplex PCR was an effective assay demonstrating high specificity for the simultaneous detection of five target pathogens in food system.

Optimal Fuzzy Sliding-Mode Control for Microcontroller-based Microfluidic Manipulation in Biochip System

  • Chung, Yung-Chiang;Wen, Bor-Jiunn
    • 제어로봇시스템학회:학술대회논문집
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    • 2004.08a
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    • pp.196-201
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    • 2004
  • In biometric and biomedical applications, a special transporting mechanism must be designed for the ${\mu}$TAS (micro total analysis system) to move samples and reagents through the microchannels that connect the unit procedure components in the system. An important issue for this miniaturization and integration is microfluid management technique, i.e., microfluid transportation, metering, and mixing. In view of this, this study presents an optimal fuzzy sliding-mode control (OFSMC) design based on the 8051 microprocessor and implementation of a complete microfluidic manipulated system implementation of biochip system with a pneumatic pumping actuator, a feedback-signal photodiodes and flowmeter. The new microfluid management technique successfully improved the efficiency of molecular biology reaction by increasing the velocity of the target nucleic acid molecules, which increases the effective collision into the probe molecules as the target molecules flow back and forth. Therefore, this hybridization chip was able to increase hybridization signal 6-fold and reduce non-specific target-probe binding and background noises within 30 minutes, as compared to conventional hybridization methods, which may take from 4 hours to overnight. In addition, the new technique was also used in DNA extraction. When serum existed in the fluid, the extraction efficiency of immobilized beads with solution flowing back and forth was 88-fold higher than that of free-beads.

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Identification and Function Prediction of Novel MicroRNAs in Laoshan Dairy Goats

  • Ji, Zhibin;Wang, Guizhi;Zhang, Chunlan;Xie, Zhijing;Liu, Zhaohua;Wang, Jianmin
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.3
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    • pp.309-315
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    • 2013
  • MicroRNAs are a class of endogenous small RNAs that play important roles in post-transcriptional gene regulation by directing degradation of mRNAs or facilitating repression of target gene translation. In this study, three small RNA cDNA libraries from the mammary gland tissues of Laoshan dairy goats (Capra hircus) were constructed and sequenced, individually. Through Solexa high-throughput sequencing and bioinformatics analysis, we obtained 50 presumptive novel miRNAs candidates, and 55,448 putative target genes were predicted. GO annotations and KEGG pathway analyses showed the majority of target genes were involved in various biological processes and metabolic pathways. Our results discovered more information about the regulation network between miRNAs and mRNAs and paved a foundation for the molecular genetics of mammary gland development in goats.

Antioxidant Activity and Its Mechanism of Paeonia lactiflora Pall Extract

  • Heo, Jee-In;Kim, Jeong-Hyeon;Lee, Jeong-Min;Kim, Sung-Chan;Park, Jae-Bong;Kim, Jaebong;Lee, Jae-Yong
    • Natural Product Sciences
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    • v.19 no.1
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    • pp.49-53
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    • 2013
  • Paeonia lactiflora Pall (PL) has been used as a traditional herbal medicine in China, Korea, and Japan for more 1,200 years. PL has reported to have antioxidant activity and protective effect of cells from oxidative stress, although the mechanism has not been verified. FOXO3a is a transcription factor that binds to its target gene's consensus FOXO binding site. FOXO3a protein modulates the various biological functions including cell cycle control, apoptosis, DNA repair, and ROS detoxification. Therefore, FOXO3a activity is associated with cancer, aging, diabetes, infertility, neurodegeneration, and immune system dysfunction. Here we found that FOXO3a was activated by PL extract. Transcriptional target genes such as MnSOD, p27, and GADD45 were activated by PL extract. Protein levels of MnSOD and catalase were increased, consequently, ROS level was reduced in HEF cells by PL extract. These findings suggest that PL extract has an antioxidant activity through FOXO activation and thereby activation of FOXO target genes, MnSOD and catalase.

BETTER UNDERSTANDING OF THE BIOLOGICAL EFFECTS OF RADIATION BY MICROSCOPIC APPROACHES

  • Kim, Eun-Hee
    • Nuclear Engineering and Technology
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    • v.40 no.7
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    • pp.551-560
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    • 2008
  • Radiation has stochastic aspects in its generation, its choice of interaction mode during traveling in media, and its impact on living bodies. In certain circumstances, like in high dose environments resulting from low-LET radiation, the variance in its impact on a target volume is negligible. On the contrary, in low dose environments, especially when they are attributed to high-LET radiation, the impact on the target carries with it a large variance. This variation is more significant for smaller target volumes. Microdosimetric techniques, which have been developed to estimate the distribution of radiation energy deposited to cellular and subcellular-sized targets, contrast with macrodosimetric techniques which count only the average value. Since cells and DNA compounds are the critical targets in human bodies, microdosimetry, or dose estimation by microscopic approach, helps one better analyze the biological effects of radiation on the human body. By utilizing microbeam systems designed for individual cell irradiation, scientists have discovered that human cells exhibit radiosensitive reactions without being hit themselves (bystander effect). During the past 10 or more years, a new therapeutic protocol using discontinuous multiple micro-slit beams has been investigated for its clinical application. It has been suggested that the beneficial bystander effect is the essence of this protocol.

Resveratrol epigenetically regulates the expression of zinc finger protein 36 in non-small cell lung cancer cell lines

  • Ahmad Fudhaili;Nal Ae Yoon;Seokmin Kang;Jinhyun Ryu;Joo Yeon Jeong;Dong Hoon Lee;Sang Soo Kang
    • Oncology Reports
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    • v.41 no.2
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    • pp.1377-1386
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    • 2019
  • Zinc finger protein 36 (ZFP36) is an AU-rich element protein that binds to 3'-untranslated regions and promotes the decay of target mRNAs. Downregulation of ZFP36 expression in turn results in stabilization of target mRNAs. A recent study indicated that downregulation of ZFP36 expression in human liver cancer is caused by epigenetic mechanisms. The purpose of the present study was to investigate the potential of resveratrol (Res) to induce ZFP36 expression. Promoter methylation was analyzed using methylation-sensitive restriction analysis. It was determined that Res treatment increased ZFP36 expression and decreased the mRNA levels of ZFP36 target genes in A549 lung cancer cells. Additionally, Res suppressed the expression of DNA (cytosine-5)-methyltransferase 1 and induced demethylation of the ZFP36 promoter. Collectively, the present results demonstrated that Res has anticancer activity through its epigenetic regulation of ZFP36 in non-small cell lung cancer.