• Title/Summary/Keyword: PCR polymorphic

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Analysis of Genetic Relationship Among Collected Cymbidium goeringii Based on RAPD (RAPD를 이용한 춘란 수집종 20 품종의 유연관계 분석)

  • Kim, Tae Bok;Lee, Jin Jae;Song, Young Ju;Choi, Chang Hak;Cheong, Dong Chun;Yu, Young Jin
    • FLOWER RESEARCH JOURNAL
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    • v.19 no.4
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    • pp.225-230
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    • 2011
  • This research was performed for making data-base of cross-breeding between Cymbidium goeringii cultivars. Morphological characteristics were investigated and then genetic relationship was analyzed. Collected 20 Cymbidium goeringii cultivars were clustered into 2 groups. Seven cultivars were clustered into group I, and thirteen cultivars were clustered into group II. Group I doesn't have leaf pattern. Group have leaf pattern. The genetic relationship among collected 20 Cymbidium goeringii cultivars was anaylzed using RAPD with ten 10-mers random primer. Eighty-nine bands were generated by RAPD. Among the rest, three bands were monomorphic and eight-six bands were polymorphic. Overall similarity degree ranged from 0.521 to 0.862. The result of RAPD analysis was clustered into 2 groups, too. Sixteen cultivars were clustered into GroupX, and four cultivars were clustered into GroupY. Result of classification with morphological characteristics and RAPD showed different pattern, but 4 cultivars of GroupY by RAPD analysis were included in groupby morphological characteristics. Crossbreeding combination among low related coltivars in RAPD analysis may get more efficient result.

Genetic variations and relationships of Phragmites japonica and P. communis according to water environment change (수환경변화에 따른 갈대와 달뿌리풀의 유전적 변이 및 유연관계)

  • Kim, Yong-Hyun;Kim, Joo-Hwan
    • Korean Journal of Plant Resources
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    • v.22 no.2
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    • pp.152-158
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    • 2009
  • We performed RAPD analysis to investigate the inter-/intraspecific relationships and regional variations of Phragmites japonica and P. communis according to the environmental change. One hundred and fourty nine genetic effective polymorphic bands between 300 bp and 1,900 bp were marked from RAPD PCR with nine oligoprimers. From the RAPD analysis by Nei-Li's genetic distance, the dissimilarity indices among the populations of Phragmites japonica were relatively low from 0.012 to 0.061, and Phragmites communis were also low from 0.033 to 0.095. It showed the close genetic relationships among the same species populations, and both species were distinctly independent with relatively high level of dissimilarity indices (0.043 - 0.132). The obvious genetic markers to distinguish two species were confirmed and those profiles were suggested. From the UPGMA phenogram by RAPD analysis, both species showed the water environment related cluster patterns by distributional regions. RAPD analysis was useful to delimit two species taxonomically and to investigate the genetic relationships among inter-/intraspecific populations.

Polymorphism analysis of tri- and tetranucleotide repeat microsatellite markers in Hanwoo cattle

  • Shil Jin;Jeong Il Won;Hyoun Ju Kim;Byoungho Park;Sung Woo Kim;Ui Hyung Kim;Sung-Sik Kang;Hyun-Jeong Lee;Sung Jin Moon;Myung Sun Park;Yong Teak Sim;Sun Sik Jang;Nam Young Kim
    • Journal of Animal Science and Technology
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    • v.66 no.4
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    • pp.717-725
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    • 2024
  • The Hanwoo traceability system currently utilizes 11 dinucleotide repeat microsatellite (MS) markers. However, dinucleotide repeat markers are known to have a high incidence of polymerase chain reaction (PCR) artifacts, such as stutter bands, which can complicate the accurate reading of alleles. In this study, we examined the polymorphisms of the 11 dinucleotide repeat MS markers currently employed in traceability systems. Additionally, we explored four trinucleotide repeat MS markers and one tetranucleotide repeat MS marker in a sample of 1,106 Hanwoo cattle. We also assessed the potential utility of the tri- and tetranucleotide repeat MS markers. The polymorphic information content (PIC) of the five tri- and tetranucleotide repeat markers ranged from 0.663 to 0.767 (mean: 0.722), sufficiently polymorphic and slightly higher than the mean (0.716) of the current 11 dinucleotide repeat markers. Using all 16 markers, the mean PIC was 0.718. The estimated probability of identity (PI) was 3.13 × 10-12 using the 11 dinucleotide repeat markers, 7.03 × 10-6 using the five tri- and tetranucleotide repeat markers, and 2.39 × 10-17 using all 16 markers; the respective PIhalf-sibs values were 2.69 × 10-9, 1.29 × 10-4, and 3.42 × 10-13; and the respective PIsibs values were 3.89 × 10-5, 9.6 × 10-3, and 3.69 × 10-7. The probability of exclusion1 (PE1) was 0.999864 for the 11 dinucleotide repeat markers, 0.981141 for five of the tri- and tetranucleotide repeat markers, and > 0.99 for all 16 markers; the respective PE2 values were 0.994632, 0.901369, and > 0.99; and the respective PE3 values were 0.998702, > 0.99, and > 0.99. The five investigated triand tetranucleotide repeat MS markers can be used in combination with the 11 existing MS markers to improve the accuracy of individual identification and paternity testing in Hanwoo.

Association Analyses with Carcass Traits in the Porcine KIAA1717 and HUMMLC2B Genes

  • Xu, D.Q.;Xiong, Y.Z.;Liu, M.;Lan, J.;Ling, X.F.;Deng, C.Y.;Jiang, S.W.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.11
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    • pp.1519-1523
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    • 2005
  • By screening a subtracted cDNA library constructed with mRNA obtained from the longissimus dorsi muscles of F1 hybrids Landrace${\times}$Yorkshire and their Yorkshire female parents, we isolated two partial sequences coding for the H3-K4-specific methyltransferase (KIAA1717) and skeletal muscle myosin regulatory light chain (HUMMLC2B) genes. In the present work we investigated two SNPs, one (C1354T) at the 3' untranslated region (UTR) of KIAA1717 and one (A345G) at the SINE (PRE-1) element of HUMMLC2B, in a resource population derived from crossing Chinese Meishan and Large White pig. The selected pigs were genotyped by means of a PCR-RFLP protocol. Significant associations were observed for the KIAA1717 C1354T polymorphic site with thorax-waist backfat thickness (p<0.05), buttock backfat thickness (p<0.05), average backfat thickness (p<0.05), loin eye height (p<0.05), loin eye area (p<0.05), carcass length to 1$^{st}$ spondyle (p<0.01) and carcass length to 1st rib (p<0.01). HUMMLC2B A345G were significantly associated with loin eye width (p<0.05), loin eye area (p<0.05). Further studies are needed to confirm these preliminary results.

Polymorphism in the promoter region of the plasminogen activator inhibitor-1 (PAI-1) gene in Kawasaki disease (가와사키병 환아에서 plasminogen activator inhibitor-1 유전자 다형성에 관한 연구)

  • Han, Mi Young
    • Clinical and Experimental Pediatrics
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    • v.50 no.6
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    • pp.570-575
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    • 2007
  • Purpose : To demonstrate genetic background of pathogenesis of Kawasaki disease (KD), I examined the genetic polymorphism of plasminogen activator inhibitor-1 (PAI-1) in KD patients. Methods : PCR-RFLP of PAI- 1 promotor gene was analyzed in 56 KD patients admitted to Kyunghee University Hospital, Gachon Medical School Gil Hospital, and Eulji Hospital from March to August 2000 and 206 normal control populations. Results : There were no differences in the genotype and allelic frequency of the PAI-1-675 (4G/5G) and PAI-1-844 (G/A) polymorphic site (which are located in the promoter region) between KD and control subjects. Also I could not detect any significant differences in specific genotypes between patients with the coronary artery lesion (CAL) and patients without CAL. Conclusion : No association was observed in -844 G/A and -675 4G/5G of PAI-1 gene polymorphism with KD.

Genetic Variability and Geographical Distribution of Mycotoxigenic Fusarium verticillioides Strains Isolated from Maize Fields in Texas

  • Ortiz, Carlos S.;Richards, Casey;Terry, Ashlee;Parra, Joselyn;Shim, Won-Bo
    • The Plant Pathology Journal
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    • v.31 no.3
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    • pp.203-211
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    • 2015
  • Maize is the dominant cereal crop produced in the US. One of the main fungal pathogens of maize is Fusarium verticillioides, the causative agent of ear and stalk rots. Significantly, the fungus produces a group of mycotoxins - fumonisins - on infested kernels, which have been linked to various illnesses in humans and animals. Nonetheless, durable resistance against F. verticillioides in maize is not currently available. In Texas, over 2.1 million acres of maize are vulnerable to fumonisin contamination, but understanding of the distribution of toxigenic F. verticillioides in maize-producing areas is currently lacking. Our goal was to investigate the genetic variability of F. verticillioides in Texas with an emphasis on fumonisin trait and geographical distribution. A total of 164 F. verticillioides cultures were isolated from 65 maize-producing counties. DNA from each isolate was extracted and analyzed by PCR for the presence of FUM1- a key fumonisin biosynthesis gene - and mating type genes. Results showed that all isolates are in fact F. verticillioides capable of producing fumonisins with a 1:1 mating-type gene ratio in the population. To further study the genetic diversity of the population, isolates were analyzed using RAPD fingerprinting. Polymorphic markers were identified and the analysis showed no clear correlation between the RAPD profile of the isolates and their corresponding geographical origin. Our data suggest the toxigenic F. verticillioides population in Texas is widely distributed wherever maize is grown. We also hypothesize that the population is fluid, with active movement and genetic recombination occurring in the field.

Genetic Relationships of Panax Species by RAPD and ISSR Analyses

  • In, Dong-Su;Kim, Young-Chang;Bang, Kyong-Hwan;Chung, Jong-Wook;Kim, Ok-Tae;Hyun, Dong-Yoon;Cha, Seon-Woo;Kim, Tae-Soo;Seong, Nak-Sul
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.5
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    • pp.249-253
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    • 2005
  • This study was carried out to develop convenient and reproducible methods for identifying the genetic relationship among germplasms of Panax species based on molecular genetics. Using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses, genetic polymorphism of the Panax species was investigated with following cultivars and accessions, such as Chunpoong, Yunpoong, Kopoong, Sunpoong, and Kumpoong in domestic cultivars, Hwangsuk, Jakyung and Suckju in domestic accessions, and Panax quinquefolius L. and Panax japonicus C.A. Meyer in foreign introduced accessions, respectively. Specific DNA fragments ranging from 200 to 3,000 base pairs in size could be obtained with various ISSR and RAPD primers under the optimized PCR conditions. The dissimilarity coefficients among the genetic polymorphisms of ginseng cultivars and accessions were calculated from 0.26 to 0.90 in RAPD and from 0.12 to 0.89 in ISSR analysis, respectively. Eleven plant samples were grouped siblings together with cultivars and parents based on cluster analysis of genetic distance depending on genetic property such as origin of the species. In results, both RAPD and ISSR analyses were useful for identifying the genetic relationship among cultivars and accessions of Panax species at DNA level.

DNA Polymorphism in SLC11A1 Gene and its Association with Brucellosis Resistance in Indian Zebu (Bos indicus) and Crossbred (Bos indicus×Bos taurus) Cattle

  • Kumar, Nishant;Ganguly, Indrajit;Singh, Rajendra;Deb, Sitangsu M.;Kumar, Subodh;Sharma, Arjava;Mitra, Abhijit
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.7
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    • pp.898-904
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    • 2011
  • The PCR- restriction fragment length polymorphism (RFLP) in and around TM4 of SLC11A1 gene and its association with the incidences of brucellosis in Hariana breed (Bos indicus) and Holstein Friesian crossbred (Bos indicus${\times}$Bos taurus) cattle was examined. A fragment of 954 bp encoding the TM4 was amplified, and RFLP was identified by digestion of the amplicon independently with AluI and TaqI. The amplicon (GenBank Acc. No. AY338470 and AY338471) comprised of a part of exon V (<59 bp) and VII (62>), and entire intron 5 (423 bp), exon VI (71 bp) and intron 6 (339 bp). Digestion with AluI revealed the presence of two alleles viz, A (281, 255, 79 and 51 bp) and B (541, 255, 79 and 51 bp). The frequency of A allele was estimated as 0.80 and 0.73 in Hariana and crossbred cattle, respectively. Due to presence of a polymorphic TaqI site at intron 5, two alleles: T (552 and 402 bp) and Q (231, 321 and 402 bp) were identified. The frequency of T allele was estimated as 0.96 and 0.97, respectively. For association study, on the basis of serological tests and history of abortion, the animals were grouped into "affected" and "non-affected". However, no association could be established with the observed RFLPs.

Polymorphic Diversity of UBX Domain D from cDNA Isolated from Pectoral Muscle of Korean Native Chicken

  • Sun, Sang-Soo;Kamyab, Abdolreza;Firman, Jeff
    • Korean Journal of Poultry Science
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    • v.38 no.3
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    • pp.191-195
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    • 2011
  • The objectives of this study are to identify specific functional genes which are related with growth and protein structure of the pectoral muscle of Korean native chicken. Pectoral muscle was isolated from three Korean native chickens (KNC, red brown, 12 months old, 2.41 ${\pm}$ 0.24 kg) and three Cornish chickens (16 month old, 2.76 ${\pm}$ 3.0 kg). The subtraction cDNA library was prepared in PCR4 Blunt-TOPO vector. The DNA sequence homology was compared with other breeds and species in GenBank. A clone NDS-81 was found to be unique for the DNA sequence homology with UBX family. Their partial sequence has high homology (98%) with chicken UBX domain D. Chicken UBX domain has chicken (93%), cattle (68%), dog (67%), mouse (64%) and, human (63%) nucleotide sequence homology. Several regions were mutated from T in chicken to C or G in the NDS-81 clone. The first site is LAD in chicken, but it was expressed as (L)RM in clone NDS-81. In this site, amino acids were changed from Ala to Arg, and from Asp to Met. The second site was changed from ER (Arg) in chicken to ED (Asp) in clone NDS-81. They are both containing functional side chains and play an important role in binding other proteins. Therefore, the clone NDS-81 could be a different candidate gene for the UBX family gene and could related with pectoral muscle structure of Korean native chicken.

Unraveling Haplotype Diversity of the Apical Membrane Antigen-1 Gene in Plasmodium falciparum Populations in Thailand

  • Lumkul, Lalita;Sawaswong, Vorthon;Simpalipan, Phumin;Kaewthamasorn, Morakot;Harnyuttanakorn, Pongchai;Pattaradilokrat, Sittiporn
    • Parasites, Hosts and Diseases
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    • v.56 no.2
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    • pp.153-165
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    • 2018
  • Development of an effective vaccine is critically needed for the prevention of malaria. One of the key antigens for malaria vaccines is the apical membrane antigen 1 (AMA-1) of the human malaria parasite Plasmodium falciparum, the surface protein for erythrocyte invasion of the parasite. The gene encoding AMA-1 has been sequenced from populations of P. falciparum worldwide, but the haplotype diversity of the gene in P. falciparum populations in the Greater Mekong Subregion (GMS), including Thailand, remains to be characterized. In the present study, the AMA-1 gene was PCR amplified and sequenced from the genomic DNA of 65 P. falciparum isolates from 5 endemic areas in Thailand. The nearly full-length 1,848 nucleotide sequence of AMA-1 was subjected to molecular analyses, including nucleotide sequence diversity, haplotype diversity and deduced amino acid sequence diversity and neutrality tests. Phylogenetic analysis and pair-wise population differentiation ($F_{st}$ indices) were performed to infer the population structure. The analyses identified 60 single nucleotide polymorphic loci, predominately located in domain I of AMA-1. A total of 31 unique AMA-1 haplotypes were identified, which included 11 novel ones. The phylogenetic tree of the AMA-1 haplotypes revealed multiple clades of AMA-1, each of which contained parasites of multiple geographical origins, consistent with the $F_{st}$ indices indicating genetic homogeneity or gene flow among geographically distinct populations of P. falciparum in Thailand's borders with Myanmar, Laos and Cambodia. In summary, the study revealed novel haplotypes and population structure needed for the further advancement of AMA-1-based malaria vaccines in the GMS.