• Title/Summary/Keyword: GENE FLOW

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Characterization of a Novel Gene in the Extended MHC Region of Mouse, NG29/Cd320, a Homolog of the Human CD320

  • Park, Hyo-Jin;Kim, Ji-Yeon;Jung, Kyung-In;Kim, Tae-Jin
    • IMMUNE NETWORK
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    • v.9 no.4
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    • pp.138-146
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    • 2009
  • Background: The MHC region of the chromosome contains a lot of genes involved in immune responses. Here we have investigated the mouse NG29/Cd320 gene in the centrometrically extended MHC region of chromosome 17. Methods: We cloned the NG29 gene by RT-PCR and confirmed the tissue distribution of its gene expression by northern blot hybridization. We generated the NG29 gene expression constructs and polyclonal antibody against the NG29 protein to perform the immunofluorescence, immunoprecipitation and flow cytometric analysis. Results: The murine NG29 gene and its human homologue, the CD320/8D6 gene, were similar in the gene structure and tissue expression patterns. We cloned the NG29 gene and confirmed its expression in plasma membrane and intracellular compartments by transfecting its expresssion constructs into HEK 293T cells. The immunoprecipitation studies with rabbit polyclonal antibody raised against the NG29-NusA fusion protein indicated that NG29 protein was a glycoprotein of about 45 kDa size. A flow cytometric analysis also showed the NG29 expression on the surface of Raw 264.7 macrophage cell line. Conclusion: These findings suggested that NG29 gene in mouse extended MHC class II region was the orthologue of human CD320 gene even though human CD320/8D6 gene was located in non-MHC region, chromosome 19p13.

Utilization of the bar gene to develop an efficient method for detection of the pollen-mediated gene flow in Chinese cabbage (Brassica rapa spp. pekinensis)

  • Lim, Chaewan;Kim, Sunggil;Choi, Yeonok;Park, Young-doo;Kim, Sung Uk;Sung, Soon-Kee
    • Plant Biotechnology Reports
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    • v.1 no.1
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    • pp.19-25
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    • 2007
  • To develop an efficient screening method for detection of the transgene in Chinese cabbage (Brassica rapa spp. pekinensis) utilizing Basta spray, optimal conditions for Basta application were examined in this study. Two transgenic Chinese cabbage lines were obtained through Agrobacterium-mediated transformation and used as transgenic positive controls in the Basta screening experiment. Differential concentrations of glufosinate-ammonium were sprayed into three different growth stages of 12 commercial Chinese cabbage cultivars. The results showed that no plants could survive higher than 0.05% glufosinate-ammonium, and plants at the 2-3 leaf stage were most vulnerable to glufosinate-ammonium. On the other hand, no damage was observed in the transgenic control plants. Reliability of the Basta spray method was proven by showing perfect co-segregation of the tolerance to glufosinate-ammonium and the presence of the bar gene in T1 segregating populations of the transgenic lines, as revealed by both PCR and Southern blot analyses. Using the developed Basta screening method, we tried to investigate the transgene flow through pollen dispersal, but failed to detect any transgene-containing non-transgenic Chinese cabbages whose parents had been planted adjacent to transgenic Chinese cabbages in field conditions. However, the transgene was successfully detected using Basta spray from the non-transgenic plants bearing the transgene introduced by hand-pollination. Since the Basta spray method developed in this study is easy to apply and economical, it will be a valuable tool for understanding the mechanism of gene flow through pollen transfer and for establishing a biosafety test protocol for genetically modified (GM) Chinese cabbage cultivars.

Additional mitochondrial DNA sequences from the dung beetle, Copris tripartitus (Coleoptera: Scarabaeidae), an endangered species in South Korea

  • Hwang, Eun Ju;Jeong, Su Yeon;Wang, Ah Rha;Kim, Min Jee;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.36 no.2
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    • pp.31-41
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    • 2018
  • The dung beetle, Copris tripartitus (Coleoptera: Scarabaeidae), is an endangered insect in South Korea. Previously, partial mitochondrial COI and CytB gene sequences have been used to infer genetic diversity and gene flow of this species in South Korea. In this study, we additionally collected C. tripartitus (n = 35) from one previous locality and two new localities, sequenced COI and CytB genes, and combined these with preexisting data for population genetic analysis. Sequence divergence of current samples showed slightly lower values [4.86% (32 bp) for COI and 4.16% (18 bp) for CytB] than that in the previous study. Nucleotide diversity (${\pi}$) ranged from 0.005336 (Gulupdo) to 0.020756 (Seogwi-dong) in COI and 0.009060 (Aewol-eup) to 0.017464 (Seogwi-dong) in CytB. Seogwi-dong samples that showed the highest ${\pi}$ in the previous study also showed the highest ${\pi}$ in this study for both gene sequences. The newly investigated Gulupdo samples had the lowest haplotype diversity for both gene sequences. They also had the lowest ${\pi}$ for COI and the second lowest ${\pi}$ for CytB. On the other hand, the newly added Haean-dong sample had relatively higher diversity estimates. Gene flow among populations was high, although significant difference was only detected between Gulupdo and Anmado or between Gulupdo and Seogwi-dong for COI sequences (P < 0.05). Considering the high genetic diversity and gene flow in C. tripartitus populations, one major issue regarding conservation seems not to be recovery of genetic diversity.

Population Genetic Structure of Octopus minor Sasaki from Korea and China Based on a Partial Sequencing of Mitochondrial 16S rRNA (미토콘드리아 16S rRNA 염기서열에 의한 한국, 중국 낙지의 유전자 집단 분석)

  • Kim, Joo-Il;Oh, Taeg-Yun;Seo, Young-Il;Cho, Eun-Seob
    • Journal of Life Science
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    • v.19 no.6
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    • pp.711-719
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    • 2009
  • We determined a portion of mitochondrial 16S rRNA gene sequences (416 bp) to investigate the genetic structure of the octopus (Octopus minor Ssaki) population in Korea and China. Samples were obtained from Korea (Yeosu, Namhae, Jindo, Muan, Geomundo and Seosan) and China (Sandong) during the period of August 2006 to September 2007. Sequence analyses of 28 individual specimens collected from 7 localities revealed 11 haplotypes, ranging in a sequence divergence of 0.2% - 1.2%. Phylogenetic analyses using PHYLIP and networks subdivided the octopus into two clades (termed clade A and B) and the nucleotide divergence between them was 0.4%. This haplotype subdivision was in accordance with geographic separation: one at Yeosu, Namhae, Muan and Jindo, and the other at Seosan, Geomundo and Sandong. On the basis of hierarchial genetic analysis, genetic distance between localities in Korea and China were also found, but a significant population differentiation was not shown in this study (p>0.05). Consequently, most of the octopus populations in Korea had considerable distribution due to the mitochondrial gene flow that resulted in a formation of a genetically homogenous structure, whereas some of the Korean and Chinese populations had different genetic structures. Gene flow among populations may be restricted due to impassable geographic barriers that promote genetic differentiation.

Haplotype Diversity and Gene Flow of the Diamondback Moth, Plutella xylostella(L.) (Lepidoptera: Yponomeutidae), in Korea (배추좀나방(나비목: 집나방과)의 haplotype 다양성과 유전자 이동률)

  • 김익수;배진식;최광호;진병래;이경로;손흥대
    • Korean journal of applied entomology
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    • v.39 no.1
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    • pp.43-52
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    • 2000
  • A portion of mitochondria1 COI gene (438 bp) was sequenced from the sampls of Plutella xylostella from four localities in Korea to investigate the population genetic structure and characteristics by measuring the magnitude of genetic diversity and the degree of gene flow among populations. Thirteen haplotypes ranging in nucleotide divergence 0.3% to 1.4%, were obtained from 21 individuals. The nucleotide divergence was similar to the other related studies, but haplotype diversity was substantially higher (mean h = 0.81). The genetic distance among geographically remote Cheju Island population and the two Kimhae populations, distant 1 lkm to each other, was not statistically significant (p<0.05). Instead, a substantial or high female gene flow was detected (Nm = 2-30). One Hawaiian haplotype of the diamondback moth obtained through GenBank search also was genetically similar to the ones obtained from this study. Collectively, the genetic population structure of the diamondback moth in Korea can be characterized into two aspects. First, the diamondback moths in Korea possesses overall moderate genetic divergence based on a high number of haplotypes. Second, a high haplotype diversity within each population due to the long distance dispersal with a substantial dispersal power and the resultant genetic similarity among geographic populations is characteristic.

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Pollen-Mediated Gene Flow between Glufosinate Ammonium-Tolerant GM and Non-GM Rice

  • Lee, Seung-Yeob;Kim, Min-Soo;Kim, Hyo-Jin;Ahn, Jeong-Ho;Baek, So-Hyeon;Shin, Woon-Chul;Kim, Hyun-Soon
    • Journal of Plant Biotechnology
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    • v.34 no.1
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    • pp.47-53
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    • 2007
  • To assess the risk of genetically modified (GM) rice on the agricultural ecosystem, agronomic characteristics, pollen longevity and outcrossing rate between GM (Iksan 483 and Milyang 204) and non-GM (their wild types and female parents) varieties were investigated using the bar gene as a tracer marker in paddy field. The agronomic characteristics of two GM rice were similar to their female-parents (non-GM rice) except heading date and 1,000 grain weight of Iksan 483, and they did not show a difference by the introgression of the bar gene as the genetic traits of rice varieties. Pollen viability was more than 90% just after shedding, and it was rapidly decreased below 50% at 5 minutes after shedding both GM and non-GM varieties. The Pollen longevity was lost after 30 minutes of anthesis. When the distance of gene flow from GM to non-GM rice detected to 6 m from the edge of GM rice plant, the maximum distance of pollen dispersal was 4.5m and 3.9m in Iksan 483 and Milyang 204, respectively, and that was increased in order of west, south, east, and north to the dominant wind direction, west-south. Mean outcrossing rate was very low as 0.003 and 0.001% within 1.5 m from the edge of Iksan 483 and Milyang 204, and the GM hybrids by the pollen dispersal did not detected over 4.5 m from the edge of GM rice plant. The results may help to establish the strategy which reduce the risk of pollen-mediated gene flow between GM and non-GM rice.

Genetic Diversity and Differentiation in Remnant Populations of Bupleurum latissimum Nakai, an Endangered Endemic Plant Species to Ulleung Island, Korea

  • Ku, Youn-Bong;Oh, Hyun-Kyung;Kong, Hak-Yang;Suh, Min-Hwan;Lee, Min-Hyo;Sviatlana, Trybush;Cho, Kang-Hyun
    • Animal cells and systems
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    • v.8 no.4
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    • pp.289-294
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    • 2004
  • Bupleurum latissimum is a narrowly endemic and endangered plant, restricted to only two small populations on steep cliffs of a small island, Ulleung Island, in Korea. The genetic diversity and population differentiation in the two remnant populations of the species were investigated using RAPD (random amplified polymorphic DNA) analysis. The Neis gene diversities were 0.146 in the smaller population of 45 individuals, and 0.151 in the larger population of 61 individuals. The genetic variation was not significantly different between these two populations. Genetic diversity within populations was not low considering the very small size of populations. Analysis of molecular variance (AMOVA) revealed higher variation within populations (65.9%) than genetic differentiation between them (34.1%). B. latissimum revealed higher population differentiation than other outbreeding species. The differentiation of the populations corresponded to low gene flow (Nem = 0.482). The cluster and principal coordination analyses provide strong support for high population differentiation, showing that all individuals of the two populations have built up population-specific clusters. Although gene flow between the two populations of B. latissimum was limited, they have preserved relatively high levels of genetic variation.

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.