• Title/Summary/Keyword: primer design

Search Result 68, Processing Time 0.03 seconds

Monitoring the presence of wild boar and land mammals using environmental DNA metabarcoding - Case study in Yangpyeong-gun, Gyeonggi-do - (환경 DNA 메타바코딩을 활용한 멧돼지 및 육상 포유류 출현 모니터링 - 경기도 양평군 일대를 중심으로 -)

  • Kim, Yong-Hwan;Han, Youn-Ha;Park, Ji-Yun;Kim, Ho Gul;Cho, Soo-Hyun;Song, Young-Keun
    • Journal of the Korean Society of Environmental Restoration Technology
    • /
    • v.24 no.6
    • /
    • pp.133-144
    • /
    • 2021
  • This study aims to estimate location of land mammals habitat by analyzing spatial data and investigate how to apply environmental DNA monitoring methodology to lotic system in Yangpyeong-gun, Gyeonggi-do. Environmental DNA sampling points are selected through spatial analysis with QGIS open source program by overlaying Kernel density of wild boar(Sus scrofa), elevation, slope and land-cover map, and 81 samples are collected. After 240 mL of water was filtered in each sample, metabarcoding technique using MiMammal universal primer was applied in order to get a whole list of mammal species whose DNA particles contained in filtered water. 8 and 22 samples showed DNA of wild boar and water deer, respectively. DNA of raccoon dog, Eurasian otter, and Siberian weasel are also detected through metabarcoding analysis. This study is valuable that conducted in outdoor lotic system. The study suggests a new wildlife monitoring methodology integrating overlayed geographic data and environmental DNA.

Molecular diversity of the VP2 of Carnivore protoparvovirus 1 (CPV-2) of fecal samples from Bogotá

  • Galvis, Cristian Camilo;Jimenez-Villegas, Tatiana;Romero, Diana Patricia Reyes;Velandia, Alejandro;Taniwaki, Sueli;Silva, Sheila Oliveira de Souza;Brandao, Paulo;Santana-Clavijo, Nelson Fernando
    • Journal of Veterinary Science
    • /
    • v.23 no.1
    • /
    • pp.14.1-14.11
    • /
    • 2022
  • Background: Carnivore protoparvovirus 1, also known as canine parvovirus type 2 (CPV-2), is the main pathogen in hemorrhagic gastroenteritis in dogs, with a high mortality rate. Three subtypes (a, b, c) have been described based on VP2 residue 426, where 2a, 2b, and 2c have asparagine, aspartic acid, and glutamic acid, respectively. Objectives: This study examined the presence of CPV-2 variants in the fecal samples of dogs diagnosed with canine parvovirus in Bogotá. Methods: Fecal samples were collected from 54 puppies and young dogs (< 1 year) that tested positive for the CPV through rapid antigen test detection between 2014-2018. Molecular screening was developed for VP1 because primers 555 for VP2 do not amplify, it was necessary to design a primer set for VP2 amplification of 982 nt. All samples that were amplified were sequenced by Sanger. Phylogenetics and structural analysis was carried out, focusing on residue 426. Results: As a result 47 out of 54 samples tested positive for VP1 screening, and 34/47 samples tested positive for VP2 980 primers as subtype 2a (n = 30) or 2b (n = 4); subtype 2c was not detected. All VP2 sequences had the amino acid, T, at 440, and most Colombian sequences showed an S514A substitution, which in the structural modeling is located in an antigenic region, together with the 426 residue. Conclusions: The 2c variant was not detected, and these findings suggest that Colombian strains of CPV-2 might be under an antigenic drift.

Development of Detection Method for Niphon spinosus, Epinephelus bruneus, and Epinephelus septemfasciatus using 16S rRNA Gene (16S rRNA를 이용한 다금바리, 자바리, 능성어 판별법 개발)

  • Park, Yong-Chjun;Jung, Yong-Hyun;Kim, Mi-Ra;Shin, Joon-Ho;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Lee, Sang-Jae;Han, Sang-Bae
    • Korean Journal of Food Science and Technology
    • /
    • v.45 no.1
    • /
    • pp.1-7
    • /
    • 2013
  • Niphon spinosus, Epinephelus bruneus, and Epinephelus septemfasciatus are involved in the Perciformes Order and Serranidae Family. When E. bruneus and E. septemfasciatus are fully grown, the striped pattern on the body gradually disappears. Therefore, morphological classification of adult fishes is quite difficult to identify the differences to N. spinosus. In this study, we investigate the method to differentiate those using PCR. To design the primers, 16S rRNA region of N. spinosus, E. bruneus, and E. septemfasciatus registered in the GeneBank (www.ncbi.nlm.nih.gov) have been used and for the analysis, Bio Edit ver. 7.0.9.0 was used. As a result, it was design NS-003-F/NS-005-R (136 bp), EB-001-F/EB-002-R (181 bp), and ES-001-F/ES-001-R (123 bp) primers for the differentiation of each 3 different fishes. Therefore, the species-specific primer sets would be a useful tool for scientific and speedy differentiation against the illegal distribution for consumer protection.

Detection Method for Identification of Pueraria mirifica (Thai kudzu) in Processed Foods (가공식품 중 태국칡(Pueraria mirifica) 혼입 판별법 개발)

  • Park, Yong-Chjun;Jin, Sang-Wook;Kim, Mi-Ra;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Lee, Sang-Jae;Han, Sang-Bae
    • Journal of Food Hygiene and Safety
    • /
    • v.27 no.4
    • /
    • pp.466-472
    • /
    • 2012
  • In this study, ribulose bisphosphate carboxylase (rbcL), RNApolymeraseC (rpoC1), intergenic spacer (psbA-trnH), and second internal transcribed spacer (ITS2) as identification markers for discrimination of P. mirifica in foods were selected. To be primer design, we obtained 719 bp, 520 bp, 348 bp, and 507 bp amplicon using universal primers from selected regions of P. mirifica. The regions of rbcL, rpoC1, and psbA-trnH were not proper for design primers because of high homology about P. mirifica, P. lobata, and B. superba. But, we had designed 4 pairs of oligonucleotide primers from ITS2 gene. Predicted amplicon from P. mirifica were obtained 137 bp and 216 bp using finally designed primers SFI12-miri-6F/SFI12-miri-7R and SFI12-miri-6F/SFI12-miri-8R, respectively. The species-specific primers distinguished P. mirifica from related species were able to apply food materials and processed foods. The developed PCR method would be applicable to food safety management for illegally distributed products in markets and internet shopping malls.

Single Nucleotide Polymorphism Exploring the 5'-Regions of Estrogen Receptor-${\alpha}$ Gene and Association With Reproduction Performance and Milk Yield in Hanwoo and Holstein Dairy Cattle (Estrogen Receptor-${\alpha}$ 유전자 5' 영역의 Single Nucleotide Polymorphism의 탐색과 한우와 Holstein에서 번식능력 및 산유능력과의 관계)

  • Yeom, Gyu-Tae;Jeon, Hyang-A;Park, Hae-Geum;Kim, Young Sin;Kim, Hyun;Kim, Jae Hwan;Seong, Hwan-Hoo;Cho, Young Moo;Cho, Jae-Hyeon;Ko, Yeoung-Gyu
    • Reproductive and Developmental Biology
    • /
    • v.38 no.3
    • /
    • pp.123-127
    • /
    • 2014
  • This study was conducted for SNPs in the 5'-regions of estrogen receptor-${\alpha}$ (ESR-${\alpha}$), and association with calving interval (CI), service per conception (SPC) and 305 days milk yield in Hanwoo and Holstein dairy cattle. The genetic improvement was incurred low reproduction performance. The objective of this study was to investigate connections between single nucleotide polymorphisms (SNP) of Estrogen receptor-${\alpha}$ (ESR-${\alpha}$) with reproduction performance (calving interval, service per conception, and 305 d milk yield) in Hanwoo and Holstein dairy cattle. Hanwoo and Holstein blood samples were collected from 183 and 124 dam of breeding farms and DNA was extracted. Primer design was based on NCBI GenBank (Accession No. AY340579). The PCR-RFLP method with Bgl I was used to genotype the cattle. The result showed two variants of the ESR-${\alpha}$ gene. The Bgl I cut the 492 bp amplification product into 322 bp and 170 bp fragments for allele G, while allele A remained uncut, resulting in two restriction fragments for homozygote G/G and three fragments for heterozygote A/G. We found two of different genotypes in these breeds, A/G and G/G. In Hanwoo, the A/G genotype frequency was 0.13, and G/G was 0.87. The CI of A/G was $382.18{\pm}10.03$ days, and G/G was $381.69{\pm}5.22$ days. The SPC of A/G was $1.62{\pm}0.16$, and G/G was $1.32{\pm}0.04$. While CI showed no significance difference, SPC exhibited significant difference (p<0.05). In Holstein cattle, the frequency of genotype A/G was 0.02 and G/G was 0.98. The 305 days milk yield of A/G was $7,253.00{\pm}936.00kg$ and of G/G was $8,747.51{\pm}204.88kg$, showing no significant difference.

Micro-silica Mixed Aqua-epoxy for Concrete Module Connection in Water : Part 1 - Material Development and Evaluation (해상 프리캐스트 콘크리트 부유체 모듈 가접합을 위한 마이크로 실리카 혼입 수중용 에폭시 접합 성능 검토 : Part 1 - 재료 개발 및 성능 검토)

  • Choi, Jin-Won;Kim, Young-Jun;You, Young-Jun;Kwon, Seung-Jun;Kim, Jang-Ho Jay
    • Journal of the Korea Concrete Institute
    • /
    • v.27 no.1
    • /
    • pp.21-28
    • /
    • 2015
  • Recent studies on concrete floating structure development focused on connection system of concrete modules. Precast concrete modules are designed to be attached by prestressing in the water, exposing the structure to the loads from water and making the construction difficult. Therefore, a development of bond material became a key issue in successful connection of floating concrete modules. In this study, micro-silica mixed aqua epoxy (MSAE) is developed for the task. Existing primer aqua epoxy, originally used as a bond material for the retrofit of concrete structures using fiber reinforced polymers, is evaluated to find the optimum micro-silica added mix proportion. Micro-silica of 0~4 volume % was mixed in standard mixture of aqua epoxy. Then, the material property tests were performed to study the effect of micro-silica in aqua epoxy by controlling the epoxy silane proportion by 0, ${\pm}5$, ${\pm}10%$. The optimum mix design of MSAE was derived based on the test results. The MSAE was used to connect concrete module specimens with the epoxy thickness variation of 5, 10, and 20mm. Then, 3-point loading test was performed to verify the bond capacity of MSAE. The results show that MSAE improves the bond capacity of concrete module.

Isolation of an Rx homolog from C. annuum and the evolution of Rx genes in the Solanaceae family

  • Shi, Jinxia;Yeom, Seon-In;Kang, Won-Hee;Park, Min-Kyu;Choi, Do-Il;Kwon, Jin-Kyung;Han, Jung-Heon;Lee, Heung-Ryul;Kim, Byung-Dong;Kang, Byoung-Cheorl
    • Plant Biotechnology Reports
    • /
    • v.5 no.4
    • /
    • pp.331-344
    • /
    • 2011
  • The well-conserved NBS domain of resistance (R) genes cloned from many plants allows the use of a PCR-based approach to isolate resistance gene analogs (RGAs). In this study, we isolated an RGA (CapRGC) from Capsicum annuum "CM334" using a PCR-based approach. This sequence encodes a protein with very high similarity to Rx genes, the Potato Virus X (PVX) R genes from potato. An evolutionary analysis of the CapRGC gene and its homologs retrieved by an extensive search of a Solanaceae database provided evidence that Rx-like genes (eight ESTs or genes that show very high similarity to Rx) appear to have diverged from R1 [an NBS-LRR R gene against late blight (Phytophthora infestans) from potato]-like genes. Structural comparison of the NBS domains of all the homologs in Solanaceae revealed that one novel motif, 14, is specific to the Rx-like genes, and also indicated that several other novel motifs are characteristic of the R1-like genes. Our results suggest that Rx-like genes are ancient but conserved. Furthermore, the novel conserved motifs can provide a basis for biochemical structural. function analysis and be used for degenerate primer design for the isolation of Rx-like sequences in other plant species. Comparative mapping study revealed that the position of CapRGC is syntenic to the locations of Rx and its homolog genes in the potato and tomato, but cosegregation analysis showed that CapRGC may not be the R gene against PVX in pepper. Our results confirm previous observations that the specificity of R genes is not conserved, while the structure and function of R genes are conserved. It appears that CapRGC may function as a resistance gene to another pathogen, such as the nematode to which the structure of CapRGC is most similar.

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
    • /
    • v.30 no.4
    • /
    • pp.347-356
    • /
    • 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.