• Title/Summary/Keyword: Gene flow

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Enhancement of Emulsion-mediated Gene Expression by Using Chitosan as a Pre-Condensing Agent

  • Chun, Soo-Kyung;Choi, Sung-Hee;Kim, Adele;Ahn, Woong-Shick;Kim, Chong-Kook
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.426.3-427
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    • 2002
  • The aim of this study was to enhance the transfection efficiency of emulsion-mediated gene expression by using chitosan, Conventional DNA/emulsion complexes and precondensed DNA/emulsion complexes were prepared by adding either naked or precondensed plasmids to cationic emulsion. The zeta potential. TEM, and size of transfection complexes were measured. In vitro transfection efficiency for boty complexes was also studied by several methods: flow cytometer, expression analysis by confocal microscope, RT-PCR, and in addition. (omitted)

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Single-cell Electroporation and Gene Transfection using MEMS-based Microdevice with Cantilever-type Microelectrode (멤스 기반의 캔틸레버 형 전극을 가진 마이크로 디바이스를 이용한 단일세포의 Electroporation 및 유전자 Transfection)

  • Cho, Young-Hak;Kim, Beom-Joon
    • Journal of the Korean Society for Precision Engineering
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    • v.27 no.5
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    • pp.85-91
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    • 2010
  • In this paper, we present details on fabrication of single-cell electroporation microdevice, practical experiments of single-cell electroporation with our fabricated microdevice. Also, the continuous electroporation for the continuous flow of cells is used for high-throughput electroporation. The delivery efficiency and cell viability tests are provided and the successful GFP transfection into cells is also evaluated with a fluorescent microscope after electroporation. This device enables to reduce the size of samples and thus the use of small amount of reagents. Also, it makes it possible to permit to avoid cell discrimination (transfected cells versus non-transfected cells) encountered when traditional bulk electroporation is held.

Agrobacterium-mediated transformation of Eleutherococcus senticosus with the squalene synthases gene derived from panax ginseng

  • Seo, Jin-Wook;Jeong, Jae-Hun;Han, Sung-Tai;Lee, Hak-Sung;Choi, Yong-Eui;Shin, Cha-Gyun
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.145.3-146
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    • 2003
  • Transgenic Eleutherococcus senticosus plants were prepared by introducing the genes for squalene synthase (SQS), hygromycin phosphotransferase (HPT) and green fluorescent Protein (GFP) through Agrobacterium-mediated transformation. The enzyme, SQS, represents a putative branch point in the isoprenoid pathway capable of diverting carbon flow specifically to the biosynthesis of phytosterol and oleanolic acid. The full SQS gene was isolated from P. ginseng roots. Early globular embryo clusters developed from embryogenic callus were used as the explant source. (omitted)

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How Do Bacteria Maximize Their Cellular Assets?

  • Kim, Juhyun
    • Microbiology and Biotechnology Letters
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    • v.49 no.4
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    • pp.478-484
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    • 2021
  • Cellular resources including transcriptional and translational machineries in bacteria are limited, yet microorganisms depend upon them to maximize cellular fitness. Bacteria have evolved strategies for using resources economically. Regulatory networks for the gene expression system enable the cell to synthesize proteins only when necessary. At the same time, regulatory interactions enable the cell to limit losses when the system cannot make a cellular profit due to fake substrates. Also, the architecture of the gene expression flow can be advantageous for clustering functionally related products, thus resulting in effective interactions among molecules. In addition, cellular systems modulate the investment of proteomes, depending upon nutrient qualities, and fast-growing cells spend more resources on the synthesis of ribosomes, whereas nonribosomal proteins are synthesized in nutrient-limited conditions. A deeper understanding of cellular mechanisms underlying the optimal allocation of cellular resources can be used for biotechnological purposes, such as designing complex genetic circuits and constructing microbial cell factories.

Influence of heading date difference on gene flow from GM to non-GM rices (GM벼에서 non-GM벼로 유전자 이동에 대한 개화기 차이의 영향 분석)

  • Oh, Sung-Dug;Chang, Ancheol;Kim, Boeun;Sohn, Soo-In;Yun, Doh-Won
    • Journal of the Korean Society of International Agriculture
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    • v.30 no.4
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    • pp.347-356
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    • 2018
  • Genetically modified (GM) crops have been increased continuously over the world and concerns about the potential risks of GM crops have also been increasing. Even though GM crops have not been cultivated commercially in Korea, it should be necessary to develop the safety assesment technology for GM crops. In this study, we investigated the influence of heading date difference on gene flow from GM to non-GM rice. In the experimental plot design, The PAC GM rice was placed in the center as a pollen donor and non-GM rice were placed in eight directions as pollen receivers. Five pollen receiver rice cultivars were Unkawng, Daebo, Saegyejinmi, Nakdong-byeo, and Ilmi which had different flowering times. A total of 266,436, 300,237, 305,223, 273,373, and 290,759 seeds were collected from Unkawng, Daebo, Saegyejinmi, Nakdong, and Ilmi, respectively, which were planted around PAC GM rice. The GM${\times}$non-GM hybrids were detected by repeated spraying of herbicide and PAT immunostrip assay. Finally, the hybrids were confirmed by PCR analysis using PAC gene specific primer. The hybrids were found in Nakdong-byeo which had the same heading date with PAC GM rice. The hybridization rate was 0.0007% at Nakdong-byeo plot. All of GM${\times}$non-GM hybrids were located within 2 m distance from the PAC GM rice zone. The physiological elements including rice heading date were found to be important factors to determine GM?rice out crossing rate with GM rice. Consideration should be taken into for many factors like the physiological elements of field heading date of rice cultivars to set up the safety management guideline for prevention of GM rice gene flow.

Mitochondrial DNA Polymorphism of the Japanese Anchovy (Engraulis japonicus Temminck & Schlegel) Collected from the Korean Offshore and Inshore Waters (한국근해 및 외해역에 채집된 멸치의 미토콘드리아 DNA 다양성)

  • Cho, Eun-Seob;Kim, Joo-Il
    • Journal of Life Science
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    • v.16 no.5
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    • pp.812-827
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    • 2006
  • To investigate the population structure and geographic distance among anchovies (Engraulis japonicus) in Korea, we compared and analyzed the mitochondrial DNA control region sequences (227 bp) of anchovies from 12 localities in inshore and offshore waters. The sequence analysis of 84 individuals showed 29 haplotypes, ranging in sequence divergence by pairwise comparisons from 0.3% to 3.5% (1 bp-12 bp). E9 haplotype of anchovies were found largely in inshore waters and also in offshore waters, which was regarded as the major source in Korean waters (58.3%). However, E26, E27, E28, and E29 haplotypes were found in westsouthern (locality 10, four among 7). Phylogenetic analysis using PHYLIP was divided into two clades (clade A and B). Most of the haplotypes, excluding E26, E27, E28, and E29, were strongly supported by bootstrap analysis (>75%), whereas the relationship between clade A and B was weakly supported by bootstrap analysis (51%). High levels of genetic diversity were found; haplotype diversity (H)=0.75-1.00, and nucleotide diversity $({\pi})=0.015-0.0244$. Analysis of $F_{ST}$ between populations in inshore waters ranged in 0.01-0.05, whereas those of offshore waters ranged in 0.01-0.58. A high gene flow occurred in inshore (Nm=22.61-34.22) and offshore (Nm=11.57-45.67) populations. The distribution of mitochondrial DNA haplotypes between westsouthern and other populations was suggestive of significantly different differentiation ($F_{ST}$=0.20-0.59, p<0.05; d=0.52, p=0.00; ${\phi}=0.02-0.41$, p<0.05). These results suggested that the overall anchovy population in the Korean peninsula caused considerable migration due to the mitochondrial gene flow between inshore and offshore populations to form a genetically homogenous and panmictic structure, although a heterogeneous population was found in this study.

Dormancy Associated Weedy Risk of the F1 Hybrid Resulted from Gene Flow from Oilseed Rape to Mustard (유채로부터 갓으로 유전자이동에 의한 교잡종의 휴면에 따른 잡초화 가능성)

  • Lim, Yeonhwa;Yook, Min-Jung;Zhang, Chuan-Jie;Nah, Gyoungju;Park, Suhyoung;Kim, Do-Soon
    • Weed & Turfgrass Science
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    • v.4 no.1
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    • pp.35-43
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    • 2015
  • To assess the dormancy associated weedy risk of the F1 hybrid generated by hybridization between Brassica juncea (maternal) and Brassica napus (paternal), seed germination, dormancy and longevity were examined sequentially after seed harvest. The F1 hybrids exhibited the intermediate characteristics of their parents in seed germination and dormancy with relatively high dormancy rate of 41.1%. In summer, F1 hybrid seeds buried in the 3 cm soil exhibited greater viability (52.4%) than those in the soil surface with greater seed longevity (74.6%) than its maternal (63.3%) and paternal (33.7%) parents at 100 days of over-summering in soil. In winter, F1 seeds buried in the soil surface were more viable than those in the 3 cm soil with greater seed longevity (83.5%) than its maternal (39.0%) and paternal (71.7%) parents at 100 days of over-wintering in soil. Therefore, it is concluded that F1 hybrid resulted from gene flow from OSR to mustard has high seed dormancy and longevity during summer and winter, suggesting its weedy risk potential. Further studies are required to examine the reproductivity and fitness cost of F1 hybrid to make a clearer conclusion of its weedy risk.

Development of a lateral flow dipstick test for the detection of 4 strains of Salmonella spp. in animal products and animal production environmental samples based on loop-mediated isothermal amplification

  • Wirawan Nuchchanart;Prapasiri Pikoolkhao;Chalermkiat Saengthongpinit
    • Animal Bioscience
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    • v.36 no.4
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    • pp.654-670
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    • 2023
  • Objective: This study aimed to develop loop-mediated isothermal amplification (LAMP) combined with lateral flow dipstick (LFD) and compare it with LAMP-AGE, polymerase chain reaction (PCR), and standard Salmonella culture as reference methods for detecting Salmonella contamination in animal products and animal production environmental samples. Methods: The SalInvA01 primer, derived from the InvA gene and designed as a new probe for LFD detection, was used in developing this study. Adjusting for optimal conditions by temperature, time, and reagent concentration includes evaluating the specificity and limit of detection. The sampling of 120 animal product samples and 350 animal production environmental samples was determined by LAMP-LFD, comparing LAMP-AGE, PCR, and the culture method. Results: Salmonella was amplified using optimal conditions for the LAMP reaction and a DNA probe for LFD at 63℃ for 60 minutes. The specificity test revealed no cross-reactivity with other microorganisms. The limit of detection of LAMP-LFD in pure culture was 3×102 CFU/mL (6 CFU/reaction) and 9.01 pg/μL in genomic DNA. The limit of detection of the LAMP-LFD using artificially inoculated in minced chicken samples with 5 hours of pre-enrichment was 3.4×104 CFU/mL (680 CFU/reaction). For 120 animal product samples, Salmonella was detected by the culture method, LAMP-LFD, LAMP-AGE, and PCR in 10/120 (8.3%). In three hundred fifty animal production environmental samples, Salmonella was detected in 91/350 (26%) by the culture method, equivalent to the detection rates of LAMP-LFD and LAMP-AGE, while PCR achieved 86/350 (24.6%). When comparing sensitivity, specificity, positive predictive value, and accuracy, LAMP-LFD showed the best results at 100%, 95.7%, 86.3%, and 96.6%, respectively. For Kappa index of LAMP-LFD, indicated nearly perfect agreement with culture method. Conclusion: The LAMP-LFD Salmonella detection, which used InvA gene, was highly specific, sensitive, and convenient for identifying Salmonella. Furthermore, this method could be used for Salmonella monitoring and primary screening in animal products and animal production environmental samples.

Intraspecific diversity and phylogeography of bony lip barb, Osteochilus vittatus, in Sundaland, as revealed by mitochondrial cytochrome oxidase I (mtCOI)

  • Imron Imron;Fajar Anggraeni;Wahyu Pamungkas;Huria Marnis;Yogi Himawan;Dessy Nurul Astuti;Flandrianto Sih Palimirmo;Otong Zenal Arifin;Jojo Subagja;Daniel Frikli Mokodongan;Rahmat Hidayat
    • Fisheries and Aquatic Sciences
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    • v.27 no.3
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    • pp.145-158
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    • 2024
  • Life history characteristics, habitat landscape, and historical events are believed to have shaped the patterns of genetic variation in many taxa. The bony lip barb, Osteohilus vittatus, represent a potamodromous fish that complete all life cycle in freshwater and is widely distributed in Southeast Asia. It usually lives in small rivers and other freshwater habitats, and movement between habitats for either food or reproduction has been typical. These life history characteristics may promote gene flow, leading to less structured populations. However, many freshwater habitats are fragmented, which restricts gene flow. We investigate how this interplay has shaped patterns of genetic variation and phylogeographic structure within this species in the Sundaland, a biodiversity hotspot with a complex geological history, using mitochondrial cytochrome oxidase I (mtCOI) as a genetic marker. Forty-six mtCOI sequences of 506 bp long were collected from ten localities, eight geographically isolated and two connected. The sequences were used for population genetic and phylogeographic analyses. Our results showed a low genetic diversity within populations but high between populations. There was a deep phylogeographic structure among geographically isolated populations but a lack of such structure in the connected habitats. Among geographically isolated populations, sequence divergence was revealed, ranging from 1.8% between Java and Sumatra populations to 12.2% between Malaysia and Vietnam. An indication of structuring was also observed among localities that are geographically closer but without connectivity. We conclude that despite high dispersal capacity, the joint effects of historical events, long-term geographic isolation associated with sea level oscillation during the Pleistocene, and restricted gene flow related to lack of habitat connectivity have shaped the phylogeographic structure within the O. vittatus over the Sundaland.

Alterations in Meningeal Blood Flow by Stimulation of Trigeminovascular System in Rats

  • Kim, Byung-Soo;Choi, Chang-Hwa;Lee, Won-Suk
    • Biomedical Science Letters
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    • v.11 no.3
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    • pp.365-373
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    • 2005
  • The aim of this study was to investigate the alterations in meningeal blood flow by stimulation of trigeminovascular system. An open cranial window was prepared on the right parietal bone of male Sprague-Dawley rats. Trigeminovascular system was stimulated by electrical stimulation of trigeminal ganglion (ETS), somatosensory (whisker) stimulation, or topical applications of capsaicin and neuropeptides including substance P and calcitonin gene-related peptide (CGRP). Neonatal capsaicin pretreatment was performed with subcutaneous administration of capsaicin (50 mg/kg) within the first 24 hours after birth. Changes in regional blood flow of dural artery (rDBF) and pial artery (rPBF) were continuously measured through the cranial window by laser-Doppler flowmetry. Both ETS and capsaicin caused a chain of alterations in rPBF and rDBF responses, i.e., an immediate transient decrease followed by rapid and marked increase in rPBF, which were significantly attenuated not only by pretreatments with L-733,060, a $NK_1$ receptor blocker, $CGRP_{8-37}$, a $CGRP_1$ receptor blocker, and 7-nitroindazole monosodium salt (7-NINA), a neuronal nitric oxide synthase inhibitor but also by neonatal capsaicin treatment. Exogenous neuropeptides including substance P and CGRP increased the meningeal blood flow, which was significantly attenuated not only by pretreatment with L-733,060 and $CGRP_{8-37}$, respectively, but also by pretreatment with 7-NINA. The rPBF response to whisker stimulation was significantly attenuated not only by trigeminovascular system injuries including nasociliary nerve denervation and neonatal capsaicin treatment but also by pretreatments with L-733,060, $CGRP_{8-37}$ and 7-NINA. These results suggest that the stimulation of trigeminovascular system causes prominent alterations in meningeal blood flow, and that neuropeptides as well as nitric oxide in the trigeminovascular system are importantly implicated in the regulation of meningeal blood flow.

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