• Title/Summary/Keyword: genotyping

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Fast and Precise: How to Measure Meiotic Crossovers in Arabidopsis

  • Kim, Heejin;Choi, Kyuha
    • Molecules and Cells
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    • v.45 no.5
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    • pp.273-283
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    • 2022
  • During meiosis, homologous chromosomes (homologs) pair and undergo genetic recombination via assembly and disassembly of the synaptonemal complex. Meiotic recombination is initiated by excess formation of DNA double-strand breaks (DSBs), among which a subset are repaired by reciprocal genetic exchange, called crossovers (COs). COs generate genetic variations across generations, profoundly affecting genetic diversity and breeding. At least one CO between homologs is essential for the first meiotic chromosome segregation, but generally only one and fewer than three inter-homolog COs occur in plants. CO frequency and distribution are biased along chromosomes, suppressed in centromeres, and controlled by pro-CO, anti-CO, and epigenetic factors. Accurate and high-throughput detection of COs is important for our understanding of CO formation and chromosome behavior. Here, we review advanced approaches that enable precise measurement of the location, frequency, and genomic landscapes of COs in plants, with a focus on Arabidopsis thaliana.

Reproductive technologies needed for the generation of precise gene-edited pigs in the pathways from laboratory to farm

  • Ching-Fu Tu;Shu-Hui Peng;Chin-kai Chuang;Chi-Hong Wong;Tien-Shuh Yang
    • Animal Bioscience
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    • v.36 no.2_spc
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    • pp.339-349
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    • 2023
  • Gene editing (GE) offers a new breeding technique (NBT) of sustainable value to animal agriculture. There are 3 GE working sites covering 5 feasible pathways to generate GE pigs along with the crucial intervals of GE/genotyping, microinjection/electroporation, induced pluripotent stem cells, somatic cell nuclear transfer, cryopreservation, and nonsurgical embryo transfer. The extension of NBT in the new era of pig breeding depends on the synergistic effect of GE and reproductive biotechnologies; the outcome relies not only on scientific due diligence and operational excellence but also on the feasibility of application on farms to improve sustainability.

An large scale single nucleotide polymorphism analysis method using mutual information and MDR (상호정보량과 MDR을 이용한 대용량 단일염기다형성 연관성 분석)

  • Jeong, Hyun-hwan;Wee, Kyubum
    • Proceedings of the Korea Information Processing Society Conference
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    • 2010.11a
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    • pp.1392-1394
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    • 2010
  • 단일염기다형성 유전형 자료에 대한 유전자형을 얻어내는 기술(genotyping)이 발달함에 따라 분석해야 하는 SNP의 개수가 수십만 개로 증가하였다. 따라서 기존의 연관성 분석(association study)연구 방법을 그대로 적용시키기는 어렵다. 본 논문에서는 상호정보량(mutual information)과 Multifactor dimensionality reduction을 이용하여 대용량의 SNP 유전형자료를 분석하는 방법을 제안하였고, 이 방법을 toluene diisocyanate-induced asthma에 대해 실험해본 결과 높은 판별력을 보이는 모델을 찾을 수 있었다.

cSNP Identification and Genotyping from C4B and BAT2 Assigned to the SLA Class III Region (돼지 SLA class III 영역 내 C4B 및 BAT2의 cSNP 동정 및 이를 이용한 유전자형 분석)

  • Kim, J.H.;Lim, H.T.;Seo, B.Y.;Lee, S.H.;Lee, J.B.;Yoo, C.K.;Jung, E.J.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.549-558
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    • 2007
  • C4B and BAT2, assigned to the SLA class III region, were recently reported on relation with human diseases. The primers for RT-PCR and RACE-PCR for CDS analysis of these genes of pig were designed by aligning the CDSs of humans and mice from GenBank. After we amplified and sequenced with these primers and cDNAs, the full-length CDSs of pig were determined. The CDS lengths of C4B and BAT2 were shown as 5226 bp and 6501 bp. In addition, the identities of nucleotide sequences with human and mouse were 76% to 87%, and the identities of amino acids were 72% to 90%. After we carried out the alignment with determined CDSs in this study and pig genomic sequences from GenBank, the primers for cSNP detection in genome were designed in intron regions that flanked one or more exons. Then, we amplified and directly sequenced with genomic DNAs of six pig breeds. Four cSNPs from C4B and three 3 cSNPs from BAT2 were identified. In addition, amino acid substitution occurred in six cSNP positions except for C4248T of C4B. By the Multiplex-ARMS method, we genotyped seven cSNPs with DNA samples used for direct sequencing. We verified that this result was the same as that analyzed using direct sequencing. To demonstrate recrudescence, we performed both direct sequencing and Multiplex-ARMS on two randomly selected DNA samples. The genotype of each sample showed the same result from both methods. Therefore, seven cSNPs were identified from C4B and BAT2 and could be used as the basic data for haplotype analysis of SLA class III region. Moreover, the Multiplex-ARMS method should be powerful for genotyping of genes assigned to the whole SLA region for the xenograft study.

Genotyping and Clinical study of Mycoplasma pneumoniae During 1996-1997 in Korea (1996년과 1997년 소아에서 유행한 Mycoplasma pneumoniae의 Genotype에 따른 임상적 연구)

  • Lim, Hong Hee;Ahn, Byung Moon;Kim, Eun Ryoung;Choi, Sug Ho;Moon, Young Ho;Kim, Il Soo
    • Pediatric Infection and Vaccine
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    • v.5 no.1
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    • pp.79-87
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    • 1998
  • Purpose : The P1 protein of Mycoplasma pneumoniae mediates the attachment of the pathogen to its host cell and elicits a strong humoral immune response during infection with this organism. Mycoplasma pneumoniae strains can be classified into two groups(I and II) by PCR method of P1 cytadhesin gene. In this study, we evaluated the prevalence, epidemiological and clinical characteristics of each group. Methods : From 155 patients with Mycoplasma pneumoniae, who admitted to the Department of Pediatrics, Sung-Ae and Kwangmyung Sung-Ae Hospital between November 1996 and October 1997, we collected their throat swabs or nasopharyngeal aspirates for DNA extraction and serum for indirect hemagglutination test of Mycoplasma pneumoniae. The group specific PCR amplification were performed using specific oligonucleotide primers designed for P1 gene genotyping. Results : Group I(137 patients, 88.4%) occurred frequently than group II(18 patients, 11.6%). In both group, the most prevalent season was winter in 1996(Nov. to Dec.) and fall in 1997(Aug. to Oct.) The prevalent age was four to six years old. The number of male was more than female in both group; Group 1(1.2:1), Goup 2(1.6:1). No significant relationship were found between two groups in duration of fever and hospital days(P>0.05). The rate of high antibody titers(>1:5120) was lower in group I(6/137, 4.4%) than group II(2/18, 11.1%). Conclusion : Group I was much more prevalent than group II during 1996~1997 in Korea. There was no difference between two groups in epidemiological and clinical parameters except the rate of high antibody titers. Further follow-up survey will be needed for the epidemiologic and clinical studies of Mycoplasma pneumoniae in Korea.

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Establishment of the High-Throughput Hair Roots' DNA Isolation System and Verification of Its Appicability for Hanwoo Traceability Using the 11 Microsatellite Makes (대량 모근 시료 DNA 분리 체계 확립과 11 microsatellite maker를 사용하는 한우 생산이력제로의 적용가능성 검증)

  • Lim, Hyun-Tae;Lee, Sang-Ho;Yoo, Chae-Kyoung;Sun, Du-Won;Cho, In-Cheol;Yoon, Du-Hak;Yang, Dae-Young;Cheong, Il-Cheong;Lee, Jung-Gyu;Jeon, Jin-Tae
    • Journal of agriculture & life science
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    • v.44 no.6
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    • pp.91-99
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    • 2010
  • We used a multiplex PCR primer set composed of 11 microsatellite (MS) markers and two sexing markers for gender detection. Genomic DNA extracted from hair roots of 3,510 Hanwoo were genotyped. Based on the 11MS markers, no animals had identical genotypes(TGLA227, BM2113, TGLA53, ETF10, SPS115, TGLA122, ETH3, ETH225, BM1824 and INRA23). The expected probability of identity among genotypes of random individuals (PI), the probability of identity among genotypes from random half-sibs ($PI_{half-sibs}$) and among genotypes of random individuals, and the probability of identity among genotypes from random sibs ($PI_{sibs}$) were estimated as $1.31{\times}10^{-23}$, $2.52{\times}10^{-16}$and $1.09{\times}10^{-6}$, respectively using the API-CALC program, version 1.0. We successfully completed the genotype analysis of 3,510 Hanwoo with a 3.93% genotyping failure rate. It was revealed that extracting DNA from the hair root was a time-efficient and cost-effective method to collect specimens for DNA isolation from live animals. This method also minimized stress for the animals during specimen collection. Among the hair roots from the back, belly, upper tail and lower tail, 5~13 hair roots of the lower tail led to the best genotype analysis results. Finally, we established a 96-well-format method of DNA preparation applicable for high- throughput genotype analysis.

Analysis of Varietal Difference and Genetic Diversity of Grapevine Cultivarsthrough the Leaf Inoculation of Colletotrichum spp. (포도 탄저병균의 엽면접종을 통한 국내 육성 포도나무의 품종 간 저항성 검정 및 유전적 다양성 분석)

  • Jang, Hyun A;Lee, Kyo-Sang;Oo, May Moe;Kwak, Tae-Seok;Yoon, Ha-Yeon;Thinn, Khaing Shwe Zin;Kim, Mi-Reu;Kim, Dae-Gyu;Lee, Jeong Jin;Lim, Gi Taek;Hur, Youn Young;Oh, Sang-Keun
    • Journal of agriculture & life science
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    • v.52 no.6
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    • pp.49-60
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    • 2018
  • Here, we reported 159 varieties of major cultivars using grapevine genetic resources to identify the resistant grape ripe rot cultivars. To do this, we performed pathogenicity assays from these grape cultivars by inoculating Colletotrichum acutatum and Colletotrichum gloeosporioides. Genotyping-by-sequencing(GBS) method was also used to compare genetic diversity among grape varieties. As a result, leaves inoculated with C. acutatum showed that 58 cultivars were susceptible, while 17 cultivars were resistant. In the case of C. gloeosporioides, 34 cultivars were found to be susceptible, while 25 cultivars were resistant. The 8 cultivars that showed resistance to both species were 'Agawan', 'Huangguan', 'Xiangfei', and 5 other cultivars from the hybrids of European and American species. Most of the varieties such as 'Emerald Seedless', 'Tano Red', and 'Rem 46-77(Aestivalis GVIT 0970)' originated in European species were identified as susceptible. These results can be used in the effective management of grape disease. In addition, these findings provide information for the development and cultivation of resistant to grape ripe rot disease cultivars.