• Title/Summary/Keyword: reference gene

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Gene Expression Analysis and Polymorphism Discovery to Investigate Drought Responsive System in Tropical Maize

  • Song, Kitae;Kim, Hyo Chul;Kim, Kyung-Hee;Moon, Jun-Cheol;Kim, Jae Yoon;Lee, Sang-Kyu;Lee, Byung-Moo
    • Plant Breeding and Biotechnology
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    • v.6 no.4
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    • pp.354-362
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    • 2018
  • Maize has high food and industrial value, whereas has difficulties in research because of their complex and huge size genome. Nested association mapping (NAM) was constructed to better understand maize genetics. However, most studies were conducted using the reference genome B73, and only a few studies were conducted on tropical maize. Ki3, one of the founder lines of the NAM population, is a tropical maize. We analyzed the genetic characteristics of Ki3 by using RNA sequencing and bioinformatics tools for various genetic studies. As results, a total of 30,526 genes were expressed, and expression profile were constructed. A total of 1,558 genes were differentially expressed in response to drought stress, and 513 contigs of them come from de novo assemblies. In addition, high-density polymorphisms including 464,930 single nucleotide polymorphisms (SNPs), 21,872 multiple nucleotide polymorphisms (MNPs) and 93,313 insertions and deletions (InDels) were found compared to reference genome. Among them, 15.0 % of polymorphisms (87,838) were passed non-synonymous test which could alter amino acid sequences. The variants have 66,550 SNPs, 5,853 MNPs, and 14,801 InDels, also proportion of homozygous type was higher than heterozygous. These variants were found in a total of 15,643 genes. Of these genes, 637 genes were found as differentially expressed genes (DEGs) under drought stress. Our results provide a genome-wide analysis of differentially expressed genes and information of variants on expressed genes of tropical maize under drought stress. Further characterization of these changes in genetic regulation and genetic traits will be of great value for improvement of maize genetics.

Long non-coding RNAs in Sus scrofa ileum under starvation stress

  • Wang, Shu;Ma, Yi Jia;Li, Yong Shi;Ge, Xu Sheng;Lu, Chang;Cai, Chun Bo;Yang, Yang;Zhao, Yan;Liang, Guo Ming;Guo, Xiao Hong;Cao, Guo Qing;Li, Bu Gao;Gao, Peng Fei
    • Animal Bioscience
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    • v.35 no.7
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    • pp.975-988
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    • 2022
  • Objective: In this study, we aimed to identify long non-coding RNAs (lncRNAs) that play important roles in starvation stress, analyze their functions, and discover potential molecular targets to alleviate starvation stress to provide a theoretical reference for subsequent in-depth research. Methods: We generated a piglet starvation stress animal model. Nine Yorkshire weaned piglets were randomly divided into a long-term starvation stress group (starved for 72 h), short-term starvation stress group (starved for 48 h), and the control group. LncRNA libraries were constructed using high-throughput sequencing of piglet ileums. Results: We obtained 11,792 lncRNAs, among which, 2,500 lncRNAs were novel. In total, 509 differentially expressed (DE)lncRNAs were identified in this study. Target genes of DElncRNAs were predicted via cis and trans interactions, and functional and pathway analyses were performed. Gene ontology functions and Kyoto encyclopedia of genes and genomes analysis revealed that lncRNA-targeted genes mainly participated in metabolic pathways, cellular processes, immune system processes, digestive systems, and transport activities. To reveal the mechanism underlying starvation stress, the interaction network between lncRNAs and their targets was constructed based on 26 DElncRNAs and 72 DEmRNAs. We performed an interaction network analysis of 121 DElncRNA-DEmRNA pairs with a Pearson correlation coefficient greater than 0.99. Conclusion: We found that MSTRG.19894.13, MSTRG.16726.3, and MSTRG.12176.1 might play important roles in starvation stress. This study not only generated a library of enriched lncRNAs in piglets, but its outcomes also provide a strong foundation to screen key lncRNAs involved in starvation stress and a reference for subsequent in-depth research.

Development of DNA Microarray for Pathogen Detection

  • Yoo, Seung Min;Keum, Ki Chang;Yoo, So Young;Choi, Jun Yong;Chang, Kyung Hee;Yoo, Nae Choon;Yoo, Won Min;Kim, June Myung;Lee, Duke;Lee, Sang Yup
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.2
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    • pp.93-99
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    • 2004
  • Pathogens pose a significant threat to humans, animals, and plants. Consequently, a considerable effort has been devoted to developing rapid, convenient, and accurate assays for the detection of these unfavorable organisms. Recently, DNA-microarray based technology is receiving much attention as a powerful tool for pathogen detection. After the target gene is first selected for the unique identification of microorganisms, species-specific probes are designed through bioinformatic analysis of the sequences, which uses the info rmation present in the databases. DNA samples, which were obtained from reference and/or clinical isolates, are properly processed and hybridized with species-specific probes that are immobilized on the surface of the microarray for fluorescent detection. In this study, we review the methods and strategies for the development of DNA microarray for pathogen detection, with the focus on probe design.

Detection of Mendelian and Parent-of-origin Quantitative Trait Loci for Meat Quality in a Cross between Korean Native Pig and Landrace

  • Choi, B.H.;Lee, Y.M.;Alam, M.;Lee, J.H.;Kim, T.H.;Kim, K.S.;Kim, J.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.12
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    • pp.1644-1650
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    • 2011
  • This study was conducted to detect quantitative trait loci (QTL) affecting meat quality in an $F_2$ reference population of Korean native pig and Landrace crossbreds. The three-generation mapping population was generated with 411 progeny from 38 $F_2$ full-sib families, and 133 genetic markers were used to produce a sex-average map of the 17 autosomes. The data set was analyzed using least squares Mendelian and parent-of-origin interval-mapping models. Lack-of-fit tests between models were used to characterize the QTL for mode of gene expressions. A total of 10 (32) QTL were detected at the 5% genome (chromosome)-wise level for the analyzed traits. Of the 42 QTL detected, 13 QTL were classified as Mendelian, 10 as paternal, 14 as maternal, and 5 as partial expressed QTL, respectively. Among the QTL detected at 5% genome-wise level, four QTL had Mendelian mode of inheritance on SSCs 5, 10, 12, and 13 for cooking loss, drip loss, crude lipid and crude protein, respectively; two QTL maternal inheritance for pH at 24-h and shear force on SSC11; three QTL paternal inheritance for CIE b and Hunter b on SSC9 and for cooking loss on SSC15; and one QTL partial expression for crude ash on SSC13, respectively. Most of the Mendelian QTL (9 of 13) had a dominant mode of gene action, suggesting potential utilization of heterosis for genetic improvement of meat quality within the cross population via marker-assisted selection.

Padina boryana, a brown alga from the Maldives: inhibition of α-MSH-stimulated melanogenesis via the activation of ERK in B16F10 cells

  • Jayawardena, Thilina U.;Sanjeewa, K.K. Asanka;Kim, Hyun-Soo;Lee, Hyo Geun;Wang, Lei;Lee, Dae-Sung;Jeon, You-Jin
    • Fisheries and Aquatic Sciences
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    • v.23 no.3
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    • pp.8.1-8.9
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    • 2020
  • Background: The present study investigates the potent skin whitening ability of ethanol extract from the brown alga, Padina boryana (PBE) which was collected in the shores of Fulhadhoo Island, the Maldives, and its specific pathways of action. The effect of PBE which contains a rich amount of polyphenols was evaluated using B16F10 murine melanoma cells and provides insight to the underlying mechanisms with reference to the inhibition of melanin formation. Methods: Melanin synthesis and cellular tyrosinase inhibition were assessed in the α-MSH-stimulated melanocytes. Melanogenic pathway-related protein expressions were investigated via Western blotting. ERK 42/44 was particularly examined considering its involvement in the melanogenic pathway. Further, RT-qPCR techniques were involved in gene expression analysis. Results: PBE dose-dependently inhibited the cellular melanin synthesis and tyrosinase levels. Western blotting revealed the potential of PBE to downregulate microphthalmia-associated transcription factor (MITF), tyrosinase, and tyrosinase-related protein-1 and protein-2 (TRP-1 and TRP-2). Moreover, results explained the phosphorylation of ERK was sustained via PBE and hence declined the ultimate melanin synthesis. Gene expression analysis reinforced the results obtained. Conclusions: The study provides substantial evidence to express the potential of PBE to inhibit B16F10 melanoma cell melanin synthesis. Concisely, results suggest the ability of PBE to be involved in medicinal and cosmeceutical applications.

Tissue-specific gene expression analysis of silkworm (Bombyx mori) by quantitative real-time RT-PCR

  • Park, Seung-Won;Kang, Seok-Woo;Goo, Tae-Won;Kim, Seong-Ryul;Lee, Gwang-Gill;Paik, Soon-Young
    • BMB Reports
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    • v.43 no.7
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    • pp.480-484
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    • 2010
  • The Bombyx mori Microarray Database (BmMDB; http://silkworm.swu.edu.cn/microarray) provides information for tissue-specific gene expression by using the whole-genome oligonucleotide microarray in the silkworm. We analyzed the tissue-specific expression patterns in the silk gland, fat body, and midgut five days of fifth instar larvae during the development of B. mori. To verify the tissue-specific expression, analysis was conducted using quantitative Real-time RT-PCR and the highly expressed endogenous Actin RNA as an intrinsic reference. Finally, we confirmed five genes, (sw15872, sw00692, sw20990, sw05300,and sw2250), out of 18 candidates expressed in two different tissues, which was consistent with the data published by Dr. Xiang's group, thereby supporting the BmMDB. Further studies for promoter regions of candidate genes can be applied in creating transgenic silkworms as biomedical insects for use in producing biomaterials, and to serve as well-characterized models for understanding the mechanism for the genetic regulation of tissue-specific development.

Gene Expression Profiling in the Pituitary Gland of Laying Period and Ceased Period Huoyan Geese

  • Luan, Xinhong;Cao, Zhongzan;Xu, Wen;Gao, Ming;Wang, Laiyou;Zhang, Shuwei
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.7
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    • pp.921-929
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    • 2013
  • Huoyan goose is a Chinese local breed famous for its higher laying performance, but the problems of variety degeneration have emerged recently, especially a decrease in the number of eggs laid. In order to better understand the molecular mechanism that underlies egg laying in Huoyan geese, gene profiles in the pituitary gland of Huoyan geese taken during the laying period and ceased period were investigated using the suppression subtractive hybridization (SSH) method. Total RNA was extracted from pituitary glands of ceased period and laying period geese. The cDNA in the pituitary glands of ceased geese was subtracted from the cDNA in the pituitary glands of laying geese (forward subtraction); the reverse subtraction was also performed. After sequencing and annotation, a total of 30 and 24 up and down-regulated genes were obtained from the forward and reverse SSH libraries, respectively. These genes mostly related to biosynthetic process, cellular nitrogen compound metabolic process, transport, cell differentiation, cellular protein modification process, signal transduction, small molecule metabolic process. Furthermore, eleven genes were selected for further analyses by quantitative real-time PCR (qRT-PCR). The qRT-PCR results for the most part were consistent with the SSH results. Among these genes, Synaptotagmin-1 (SYT1) and Stathmin-2 (STMN2) were substantially over-expressed in laying period compared to ceased period. These results could serve as an important reference for elucidating the molecular mechanism of higher laying performance in Huoyan geese.

Specific and Sensitive Detection of Phoma glomerata Using PCR Techniques (PCR 기법을 이용한 Phoma glomerate 의 특이검출)

  • Yun, Yeo Hong;Suh, Dong Yeon;Kim, Hyun Ju;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.41 no.1
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    • pp.52-55
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    • 2013
  • Phoma glomerata (Corda) Wollenw. & Hochapfel is a pathogenic fungus causing spot diseases of plant leaves and fruits. This fungus is important in plant quarantine of seedlings and fruits in Korea. The aim of this study was to develop a sensitive and effective diagnostic method for P. glomerata detection in imported plants. The fungal species-specific PCR primers were designed based on the nucleotide sequences of the translation elongation factor 1 alpha gene and their specificity and sensitivity were tested. The designed primers named as PhoGlo-F and PhoGlo-R amplified specifically a 170 bp sized DNA band of the target gene from the genomic DNA of P. glomerata. No amplicon was produced from genomic DNAs of 16 other Phoma spp. and reference fungal species tested. Moreover, PhoGlo-F/PhoGlo-R primers successfully worked with real-time PCR technique. The detection limit of DNA content by conventional and real-time PCR were 10 pg and 1pg of the genomic DNA of P. glomerata, respectively. We believed that the developed makers would be very useful for P. glomerata detection.

Molecular Characterization of A Novel Bacillus thuringiensis Strain from China

  • Qi Xu Feng;Li Ming Shun;Choi Jae Young;Kim Yang-Su;Wang Yong;Kang Joong Nam;Choi Heekyu;Je Yeon Ho;Song Ji Zhen;Li Jian Hong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.11 no.1
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    • pp.57-61
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    • 2005
  • A strain of Bacillus thuringiensis that showed signifi­cantly high toxicity to Plutella xylostella was isolated from a dust sample collected from Chinese tobacco warehouse and characterized. The isolate named B. thuringiensis LY-99 was determined to belong to subsp. alesti (H3a3c) by an H antisera agglutination test and produced bipyramidal inclusions. Plasmid and crystal protein patterns of the LY-99 were different from those of the reference strain, subsp. alesti. PCR analysis with specific primers revealed that this isolate contained abundant cry genes including crylAa, crylAc, crylB, crylD, crylE, crylF and cry2 genes, which was absolutely different from cry gene profile of the subsp. alesti. In addition, insecticidal activity of the LY-99 against P. xylostella larvae was about 44 times higher than that of the subsp. alesti.

Development of Detection Method of Unapproved Genetically Modified Potato (EH92-527-1) in Korea using Duplex Polymerase Chain Reaction (Duplex PCR을 이용한 국내 미승인 유전자변형 감자(EH92-527-1)의 검사법 개발)

  • Yoo, Myung-Ryul;Kim, Jae-Hwan;Yea, Mi-Chi;Kim, Hae-Yeong
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.156-160
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    • 2013
  • A duplex polymerase chain reaction (PCR) method was developed to detect unapproved genetically modified (GM) potato (EH92-527-1) in Korea. The UDP-glucose pyrophosphorylase (UGP) gene was selected as an endogenous reference gene for potato and used to validate the specificity for 14 different crops. The primer pair EH92-F/R was designed to amplify the junction sequence between the genome and transgenic region introduced in GM potato. Its specificity was also validated using several different GM events. The detection limit of the duplex PCR method is approximately 0.05%. This duplex PCR method could be useful for monitoring cultivation of unauthorized GM potato in Korea.