• Title/Summary/Keyword: Genomic analysis

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A Gene Functional Study of Rice Using Ac/Ds Insertional Mutant Population

  • Kim, So-Young;Kim, Chang-Kug;Kang, Min;Ji, Seung-Uk;Yoon, Ung-Han;Kim, Yong-Hwan;Lee, Gang-Seob
    • Plant Breeding and Biotechnology
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    • v.6 no.4
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    • pp.313-320
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    • 2018
  • Rice is the staple food of more than 50% of the world population. Cultivated rice has the AA genome (diploid, 2n = 24) and small genome size of only 430 megabase (haploid genome). As the sequencing of rice genome was completed by the International Rice Genome Sequencing Project (IRGSP), many researchers in the world have been working to explore the gene function on rice genome. Insertional mutagenesis has been a powerful strategy for assessing gene function. In maize, well characterized transposable elements have traditionally been used to clone genes for which only phenotypic information is available. In rice endogenous mobile elements such as MITE and Tos have been used to generate gene-tagged populations. To date T-DNA and maize transposable element systems have been utilized as main insertional mutagens in rice. The Ac/Ds system offers the advantage of generating new mutants by secondary transposition from a single tagged gene. To enhance the efficiency of gene detection, advanced gene-tagging systems (i.e. activation, gene or enhancer trap) have been employed for functional genomic studies in rice. Internationally, there have been many projects to develop large scales of insertional mutagenized populations and databases of insertion sites has been established. Ultimate goals of these projects are to supply genetic materials and informations essential for functional analysis of rice genes and for breeding using agronomically important genes. In this report, we summarize the current status of Ac/Ds-mediated gene tagging systems that has been conducted by collaborative works in Korea.

Analysis of genetic characteristics of pig breeds using information on single nucleotide polymorphisms

  • Lee, Sang-Min;Oh, Jae-Don;Park, Kyung-Do;Do, Kyoung-Tag
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.4
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    • pp.485-493
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    • 2019
  • Objective: This study was undertaken to investigate the genetic characteristics of Berkshire (BS), Landrace (LR), and Yorkshire (YS) pig breeds raised in the Great Grandparents pig farms using the single nucleotide polymorphisms (SNP) information. Methods: A total of 25,921 common SNP genotype markers in three pig breeds were used to estimate the expected heterozygosity ($H_E$), polymorphism information content, F-statistics ($F_{ST}$), linkage disequilibrium (LD) and effective population size ($N_e$). Results: The chromosome-wise distribution of $F_{ST}$ in BS, LR, and YS populations were within the range of 0-0.36, and the average $F_{ST}$ value was estimated to be $0.07{\pm}0.06$. This result indicated some level of genetic segregation. An average LD ($r^2$) for the BS, LR, and YS breeds was estimated to be approximately 0.41. This study also found an average $N_e$ of 19.9 (BS), 31.4 (LR), and 34.1 (YS) over the last 5th generations. The effective population size for the BS, LR, and YS breeds decreased at a consistent rate from 50th to 10th generations ago. With a relatively faster $N_e$ decline rate in the past 10th generations, there exists possible evidence for intensive selection practices in pigs in the recent past. Conclusion: To develop customized chips for the genomic selection of various breeds, it is important to select and utilize SNP based on the genetic characteristics of each breed. Since the improvement efficiency of breed pigs increases sharply by the population size, it is important to increase test units for the improvement and it is desirable to establish the pig improvement network system to expand the unit of breed pig improvement through the genetic connection among breed pig farms.

Genetic diversity analysis of fourteen geese breeds based on microsatellite genotyping technique

  • Moniem, Hebatallah Abdel;Zong, Yang Yao;Abdallah, Alwasella;Chen, Guo-hong
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.11
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    • pp.1664-1672
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    • 2019
  • Objective: This study aimed to measure genetic diversity and to determine the relationships among fourteen goose breeds. Methods: Microsatellite markers were isolated from the genomic DNA of geese based on previous literature. The DNA segments, including short tandem repeats, were tested for their diversity among fourteen populations of geese. The diversity was tested on both breeds and loci level and by mean of unweighted pair group method with arithmetic mean and structure program, phylogenetic tree and population structure were tested. Results: A total of 108 distinct alleles (1%) were observed across the fourteen breeds, with 36 out of the 108 alleles (33.2%) being unique to only one breed. Genetic parameters were measured per the 14 breeds and the 9 loci. Medium to high heterozygosity was reported with high effective numbers of alleles (Ne). Polymorphic information contents (PIC) of the screened loci was found to be highly polymorphic for eleven breeds; while 3 breeds were reported moderately polymorphic. Breeding coefficient ($F_{IS}$) ranged from -0.033 to 0.358, and the pair wise genetic differentiation ($F_{ST}$) ranged from 0.01 to 0.36 across the fourteen breeds; for the 9 loci observed and expected heterozygosity, and Ne were same as the breeds parameters, PIC of the screened loci reported 6 loci highly polymorphic and 3 loci to be medium polymorphic, and $F_{IS}$ ranged from -0.113 to 0.368. In addition, genetic distance estimate revealed a close genetic distance between Canada goose and Hortobagy goose breeds by 0.04, and the highest distance was between Taihu goose and Graylag goose (anser anser) breed by 0.54. Conclusion: Cluster analyses were made, and they revealed that goose breeds had hybridized frequently, resulting in a loss of genetic distinctiveness for some breeds.

Development of molecular markers for the differentiation of Angelica gigas Jiri line by using ARMS-PCR analysis (세발당귀(Angelica gigas Jiri)의 판별을 위한 ARMS-PCR용 분자표지 개발)

  • Lee, Shin-Woo;Lee, Soo Jin;Han, Eun-Hee;Shin, Yong-Wook;Kim, Yun-Hee
    • Journal of Plant Biotechnology
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    • v.48 no.1
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    • pp.26-33
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    • 2021
  • Angelica is a widely used medicinal and perennial plant. Information on the genetic diversity of Angelica populations is essential for their conservation and germ plasmic utilization. Although Angelica is an important medicinal plant species registered in South Korea, no molecular markers are currently available to distinguish it from other similar species from different countries. This developed single nucleotide polymorphism (SNP) markers derived from nuclear ribosomal DNA internal transcribed spacer regions genomic sequences to identify distinct Korean-specific Angelica species via amplification refractory mutation system (ARMS)-PCR curve analyses. We performed molecular authentication of different kinds of Korean-specific Angelica species such as A. gigas Nakai and A. gigas Jiri using DNA sequences in the ITS intergenic region. The SNP markers developed in this study are useful for rapidly identifying specific Angelica species from different countr.

Rapid and Visual Detection of Vibrio parahaemolyticus in Aquatic Foods Using blaCARB-17 Gene-Based Loop-Mediated Isothermal Amplification with Lateral Flow Dipstick (LAMP-LFD)

  • Hu, Yuan-qing;Huang, Xian-hui;Guo, Li-qing;Shen, Zi-chen;LV, Lin-xue;Li, Feng-xia;Zhou, Zan-hu;Zhang, Dan-feng
    • Journal of Microbiology and Biotechnology
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    • v.31 no.12
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    • pp.1672-1683
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    • 2021
  • Vibrio parahaemolyticus is recognized as one of the most important foodborne pathogens responsible for gastroenteritis in humans. The blaCARB-17 gene is an intrinsic β-lactamase gene and a novel species-specific genetic marker of V. parahaemolyticus. In this study, a loop-mediated isothermal amplification (LAMP) assay combined with a lateral flow dipstick (LFD) was developed targeting this blaCARB-17 gene. The specificity of LAMP-LFD was ascertained by detecting V. parahaemolyticus ATCC 17802 and seven other non-V. parahaemolyticus strains. Finally, the practicability of LAMP-LFD was confirmed by detection with V. parahaemolyticus-contaminated samples and natural food samples. The results showed that the optimized reaction parameters of LAMP are as follows: 2.4 mmol/l Mg2+, 0.96 mmol/l dNTPs, 4.8 U Bst DNA polymerase, and an 8:1 ratio of inner primer to outer primer, at 63℃ for 40 min. The optimized reaction time of the LFD assay is 60 min. Cross-reactivity analysis with the seven non-V. parahaemolyticus strains showed that LAMP-LFD was exclusively specific for V. parahaemolyticus. The detection limit of LAMP-LFD for V. parahaemolyticus genomic DNA was 2.1 × 10-4 ng/μl, corresponding to 630 fg/reaction and displaying a sensitivity that is 100-fold higher than that of conventional PCR. LAMP-LFD in a spiking study revealed a detection limit of approximately 6 CFU/ml, which was similar with conventional PCR. The developed LAMP-LFD specifically identified the 10 V. parahaemolyticus isolates from 30 seafood samples, suggesting that this LAMP-LFD may be a suitable diagnostic method for detecting V. parahaemolyticus in aquatic foods.

Complete mitochondrial genome of Rotunda rotundapex Miyata & Kishida 1990 (Lepidoptera: Bombycidae), which was named as Bombyx shini Park & Sohn 2002

  • Park, Jeong Sun;Kim, Min Jee;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.44 no.2
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    • pp.55-64
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    • 2022
  • Bombyx shini Park & Sohn, 2002 (Lepidoptera: Bombycidae), which was listed as an endemic species in South Korea has recently been renamed as the East Asian silk moth Rotunda rotundapex Miyata & Kishida, 1990 (Lepidoptera: Bombycidae). In this study, we sequenced the complete mitochondrial genome (mitogenome) of the R. rotundapex to announce genomic characteristics and to clarify its validity with a new name. The 15,294-bp long complete mitogenome comprises a typical set of genes [13 protein-coding genes (PCGs), 2 rRNA genes, and 22 tRNA genes] and one major noncoding, A + T-rich region, with an arrangement identical to that observed in most lepidopteran mitogenomes. The A/T content of the whole mitogenome was 79.22%; however, it varied among the regions/genes as follows: A + T-rich region, 91.62%; srRNA, 84.67%; lrRNA, 83.01%; tRNAs, 81.43%; and PCGs, 77.46%. Phylogenetic analyses of 35 species in the Bombycoidea superfamily showed the sister relationship between the families Sphingidae and Bombycidae s. str., with the higher nodal support [bootstrap support (BS) = 78%]. The Saturniidae was placed as the sister to the two families, but the nodal support for this relationship was low (BS = 53%). Current R. rotundapex was placed together with previously reported con-species with the highest nodal support, forming a separate clade from Bombyx, validating that B. shini can have a new genus name, Rotunda. However, the Korean R. rotundapex showed a substantial sequence divergence at 5.28% to that originated from an individual of type locality Taiwan in 1,459-bp of COI sequences. Considering such a high sequence divergence an additional study, which includes morphological and DNA barcoding data from further extensive distributional range maybe is needed for further robust taxonomic conclusion.

Relationship between Obesity, Gingival Inflammation, and Periodontal Bacteria after 4-Week Weight Control Program in 20's

  • Seo, Min-Seock;Hwang, Soo-Jeong
    • Journal of dental hygiene science
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    • v.22 no.2
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    • pp.99-107
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    • 2022
  • Background: Obesity weakens acquired immunity and causes infection. This study aimed to investigate the relationship between the inflammatory markers in the gingival crevicular fluid and serum and periodontal bacteria in saliva through obesity control for 4 weeks. Methods: Forty-six subjects with a body mass index (BMI) of ≥23 kg/m2 stayed in the camp for 4 weeks, followed by exercise and a low salt-low fat diet. Body size measurements, oral examinations, blood, saliva, and gingival crevicular fluid were collected before and after the program. C-reactive protein (CRP) in serum, matrix metalloproteinase (MMP)-8, MMP-9, and interleukin (IL)-1β in the gingival sulcus fluid were measured. After extracting bacterial genomic DNA from saliva, the presence of periodontal bacteria were detected using Taq probe. The relationship of each index before and after the program was analyzed through paired t-test and partial correlation analysis. Results: Campylobacter rectus (Cr) increased after the program, and there was no significant change in other bacteria. Serum CRP and Fusobacterium nucleatum (Fn), Aggregatibacter actinomycetemcomitans, Cr, ratio of Fn, and ratio of Cr had a positive relationship at baseline; however, the relationship was not significant after the program. Ratio of Prevotella intermedia had a positive relationship with MMP-9, MMP-8, IL-1β at baseline. Moreover, the ratio of Treponema denticola and the ratio of Tannerella forsythia showed a positive relationship with MMP-8, MMP-9, and IL-1β. The relationship between the ratio of Porphyromonas gingivalis and IL-1β showed a constant positive relationship at baseline and after the program. Conclusion: Obesity control program in subjects with a BMI of ≥23 kg/m2 accompanied by diet and exercise did not affect the changes in periodontal bacteria itself, but changes in the relationship between periodontal bacteria and serum CRP, the relationship between the inflammatory index in the gingival crevicular fluid and periodontal bacteria was observed.

Variovorax terrae sp. nov. Isolated from Soil with Potential Antioxidant Activity

  • Woo, Chae Yung;Kim, Jaisoo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.7
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    • pp.855-861
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    • 2022
  • A white-pigmented, non-motile, gram-negative, and rod-shaped bacterium, designated CYS-02T, was isolated from soil sampled at Suwon, Gyeonggi-do, Republic of Korea. Cells were strictly aerobic, grew optimally at 20-28℃ and hydrolyzed Tween 40. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain CYS-02T formed a lineage within the family Comamonadaceae and clustered as members of the genus Variovorax. The closest members were Variovorax guangxiensis DSM 27352T (98.6% sequence similarity), Variovorax paradoxus NBRC 15149T (98.5%), and Variovorax gossypii JM-310T (98.3%). The principal respiratory quinone was Q-8 and the major polar lipids contain phosphatidylethanolamine (PE), phosphatidylethanolamine (PG), and diphosphatidylglycerol (DPG). The predominant cellular fatty acids were C16:0, summed feature 3 (C16:1ω7c and/or C16:1ω6c) and summed feature 8 (C18:1ω7c and/or C18:1ω6c). The DNA GC content was 67.7 mol%. The ANI and dDDH values between strain CYS-02T and the closest members in the genus Variovorax were ≤ 79.0 and 22.4%, respectively, and the AAI and POCP values between CYS-02T and the other related species in the family Comamonadaceae were > 70% and > 50%, respectively. The genome of strain CYS-02T showed a putative terpene biosynthetic cluster responsible for antioxidant activity which was supported by DPPH radical scavenging activity test. Based on genomic, phenotypic and chemotaxonomic analyses, strain CYS-02T was classified into a novel species in the genus Variovorax, for which the name Variovorax terrae sp. nov., has been proposed. The type strain is CYS-02T (= KACC 22656T = NBRC 00115645T).

Characterization of the first mitogenomes of the smallest fish in the world, Paedocypris progenetica, from peat swamp of Peninsular Malaysia, Selangor, and Perak

  • Hussin, NorJasmin;Azmir, Izzati Adilah;Esa, Yuzine;Ahmad, Amirrudin;Salleh, Faezah Mohd;Jahari, Puteri Nur Syahzanani;Munian, Kaviarasu;Gan, Han Ming
    • Genomics & Informatics
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    • v.20 no.1
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    • pp.12.1-12.7
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    • 2022
  • The two complete mitochondrial genomes (mitogenomes) of Paedocypris progenetica, the smallest fish in the world which belonged to the Cyprinidae family, were sequenced and assembled. The circular DNA molecules of mitogenomes P1-P. progenetica and S3-P. progenetica were 16,827 and 16,616 bp in length, respectively, and encoded 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and one control region. The gene arrangements of P. progenetica were identical to those of other Paedocypris species. BLAST and phylogenetic analyses revealed variations in the mitogenome sequences of two Paedocypris species from Perak and Selangor. The circular DNA molecule of P. progenetica yield a standard vertebrate gene arrangement and an overall nucleotide composition of A 33.0%, T 27.2%, C 23.5%, and G 15.5%. The overall AT content of this species was consistent with that of other species in other genera. The negative GC-skew and positive AT-skew of the control region in P. progenetica indicated rich genetic variability and AT nucleotide bias, respectively. The results of this study provide genomic variation information and enhance the understanding of the mitogenome of P. progenetica. They could later deliver highly valuable new insight into data for phylogenetic analysis and population genetics.

Alaria alata (Digenea: Diplostomidae) from Korean Raccoon Dog, Nyctereutes procyonoides koreensis, in Korea

  • Lee, Heon Woo;Hong, Eui Ju;Kim, Hyeon Cheol;Ryu, Si Yun;Park, Bae Keun
    • Parasites, Hosts and Diseases
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    • v.60 no.5
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    • pp.361-365
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    • 2022
  • We report a species of diplostomid fluke recovered from 3 carcasses of wild Korean raccoon dog, Nyctereutes procyonoides koreensis, in Korea. A total of 107 diplostomid flukes were recovered from the small intestines of Korean raccoon dogs, which were obtained from the Gangwon Wildlife Medical Rescue Center. Worms fixed with 10% neutral formalin were subjected to microscopic observation and those fixed in 70% ethanol were used for molecular genomic analysis. The worm was divided into 2 separate parts, forebody and hindbody, with a total length of 3,020-4,090 (3,855) ㎛ and a width of 1,210-1,770 (1,562) ㎛. The boat-shaped forebody has a pair of characteristic tentacular appendage, 2 suckers, holdfast organ, and vitelline follicles. The oval to cylindrical hindbody has reproductive organs. The ovary was round or elliptical and located in the anterior of the testes. Two large testes were slightly segmented and tandemly arranged, occupying almost half of hindbody. The short uterus contained a relatively small number of unembryonated eggs sized 130-140×85-96 ㎛. The partial sequence of 18S rRNA of this fluke was consistent with Alaria alata. Based on the morphological and molecular characteristics, the diplostomid flukes recovered from the small intestine of Korean raccoon dogs were identified as A. alata (Digenea: Diplostomidae).