• Title/Summary/Keyword: Specific loci

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Detection of single-nucleotide polymorphism in RPB2 of Wolfiporia hoelen strains and assessment of its applicability for strain breeding (복령 균주의 RPB2 유전자 내 단일염기다형성 및 육종 활용성 분석)

  • Su Yeon, Kim;Mi-Jeong, Park;Seong Hwan, Kim;Kang-Hyeon, Ka
    • Journal of Mushroom
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    • v.20 no.4
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    • pp.199-207
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    • 2022
  • The demand for novel strains has been rising in the domestic market to increase the production of sclerotia from Wolfiporia hoelen. To improve strain breeding efficiency, we investigated whether single-nucleotide polymorphisms (SNPs) in the RNA polymerase II subunit (RPB2) gene, which may be linked to the mating type locus, are useful for distinguishing monokaryons from dikaryons in Korean W. hoelen strains. We designed a specific primer set to efficiently amplify a region of RPB2 using PCR with the genomic DNA of 12 cultivated strains and 31 wild strains of W. hoelen collected from Korea. Nucleotide sequences of the PCR-amplified RPB2 genes were determined and analyzed for the presence of SNPs among the 43 W. hoelen strains. Previously reported SNP loci were detected in the RPB2 gene of all W. hoelen strains tested. However, these previously reported SNP loci could not be applied to differentiate monokaryons from dikaryons in approximately one-third of Korean wild strains with homozygous genotypes. Three additional SNPs in the RPB2 gene, which may improve the ability to distinguish monokaryons from dikaryons, were identified by searching through the multiple sequence alignments of the 43 W. hoelen strains. The applicability of these three novel SNPs, together with the previously known SNPs, in the RPB2 gene to W. hoelen strain breeding was verified by examining the hybrid strains and their parental strains.

Development of an Effective PCR Technique for Analyzing T-DNA Integration Sites in Brassica Species and Its Application (배추과에서 T-DNA 도입 위치 분석을 위한 효과적인 PCR 방법 개발 및 이용)

  • Lee, Gi-Ho;Yu, Jae-Gyeong;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.33 no.2
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    • pp.242-250
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    • 2015
  • Insertional mutagenesis induced by T-DNA or transposon tagging offers possibilities for analysis of gene function. However, its potential remains limited unless good methods for detecting the target locus are developed. We describe a PCR technique for efficient identification of DNA sequences adjacent to the inserted T-DNA in a higher plant, Chinese cabbage (Brassica rapa ssp. pekinensis). This strategy, which we named variable argument thermal asymmetric interlaced PCR (VA-TAIL PCR), was designed by modifying a single-step annealing-extension PCR by including a touch-up PCR protocol and using long gene-specific primers. Amplification efficiency of this PCR program was significantly increased by employing an autosegment extension method and linked sequence strategy in nested long gene-specific primers. For this technique, arbitrary degenerate (AD) primers specific to B. rapa were designed by analyzing the Integr8 proteome database. These primers showed higher accuracy and utility in the identification of flanking DNA sequences from individual transgenic Chinese cabbages in a large T-DNA inserted population. The VA-TAIL PCR method described in this study allows the identification of DNA regions flanking known DNA fragments. This method has potential biotechnological applications, being highly suitable for identification of target genomic loci in insertional mutagenesis screens.

Genetic Diversity and Population Structure of Korean Soybean Collection Using 75 Microsatellite Markers

  • Lee, Gi-An;Choi, Yu-Mi;Yi, Jung-Yoon;Chung, Jong-Wook;Lee, Myung-Chul;Ma, Kyung-Ho;Lee, Sok-Young;Cho, Jin-Woong;Lee, Jung-Ro
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.4
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    • pp.492-497
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    • 2014
  • Soybean (Glycine max L.) is crucial legume crop as source of high quality vegetable protein and oil, and Korea is regarded as a part of center of soybean origin. To expand the information of conserved genetic diversity, we analyzed the genetic variability of soybean collection mainly introduced Korean accessions using 75 microsatellite markers. A total of 1,503 alleles with an average value of 20.0 alleles were detected among 644 accessions. Korean collection revealed average allele number of 13.4 while Chinese, Japanese and Southeast Asian accessions showed 9.0, 5.4 and 6.5 mean alleles, respectively. Especially, Korean accessions showed more number of private allele per locus as 3.4 contrary to other geographical groups. The mean expected heterozygosity and polymorphic information content was 0.654 and 0.616, respectively, and expected heterozygosity values were not significantly distinguished according to the geographical groups. The phylogenetic dendrogram and deduced population structure based on DNA profiles of 75 SSR loci showed Korean accessions formed distinct gene pool against Chinese accessions, and could be divided into five subpopulations. Korean soybean accessions have specific genetic diversity and might be serve the valuable alleles for bio-industry as a part of the center of soybean origin.

Diet composition of the Korean wild boar Sus scrofa coreanus (Suidae) at Mt. Jeombongsan, Korea

  • Shin, Hyung-Min;Kim, Jihee;Jin, Seon Deok;Won, Ho-Yeon;Park, Sangkyu
    • Journal of Ecology and Environment
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    • v.44 no.3
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    • pp.143-154
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    • 2020
  • Background: Korean wild boars (Sus scrofa coreanus Heude), because of their adaptability, are a widespread large mammal; however, they sometimes cause problems by invading farms and eating the crops, creating insufficiencies of some foods in South Korea. To understand the diet composition of Korean wild boars according to sex and body size, we collected their feces from Mt. Jeombongsan, Seoraksan National Park, South Korea. The sizes of fecal samples were measured, and genomic DNA was extracted from the samples. We amplified specific loci targeting plants (rbcL and trnL) and animals (COI) to detect the food sources of this omnivore and amplified the ZF and SRY regions to determine the sex. Results: In the wild boar feces, Rosaceae and Bryophyte were the most frequently detected plant food sources at the family level and Diptera and Haplotaxida were the most frequently detected animal food sources at the order level. As a result of sex determination, the sex ratio of wild boars collected in the Mt. Jeombongsan area was approximately 1:1. Our result suggested that there is no significant difference between the diet composition of male and female boars. Based on the average cross-sectional area of the feces, the top 25% were classified into the large body size group and the bottom 25% were classified into the small body size group. The large body size group mainly preferred Actinidiaceae, and the small body size group most frequently consumed Fagaceae. The diet of the large body size group was more diverse than the small body size group. Conclusions: Our results showed that the wild boars preferred Rosaceae, especially Sanguisorba and Filipendula, as plant food sources, and Diptera and Coleoptera of Insecta as animal food sources. Based on the results, the dietary preferences of wild boar appear to be distinguished by not their sex but their body size. Our study could help to elucidate the feeding ecology and population structure of wild boar, as well as address conservation and management issues.

Y-Single Nucleotide Polymorphisms Diversity in Chinese Indigenous Horse

  • Han, Haoyuan;Zhang, Qin;Gao, Kexin;Yue, Xiangpeng;Zhang, Tao;Dang, Ruihua;Lan, Xianyong;Chen, Hong;Lei, Chuzhao
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.8
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    • pp.1066-1074
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    • 2015
  • In contrast to high genetic diversity of mitochondrial DNA (mtDNA), equine Y chromosome shows extremely low variability, implying limited patrilines in the domesticated horse. In this study, we applied direct sequencing and restriction fragment length polymorphism (RFLP) methods to investigate the polymorphisms of 33 Y chromosome specific loci in 304 Chinese indigenous horses from 13 breeds. Consequently, two Y-single nucleotide polymorphisms (SNPs) (Y-45701/997 and Y-50869) and one Y-indel (Y-45288) were identified. Of those, the Y-50869 (T>A) revealed the highest variation frequency (24.67%), whereas it was only 3.29% and 1.97% in Y-45288 (T/-) and Y-45701/997 (G>T) locus, respectively. These three mutations accounted for 27.96% of the total samples and identified five Y-SNP haplotypes, demonstrating genetic diversity of Y chromosome in Chinese horses. In addition, all the five YSNP haplotypes were shared by different breeds. Among 13 horse breeds analyzed, Balikun horse displayed the highest nucleotide diversity (${\pi}=5.6{\times}10^{-4}$) and haplotype diversity (h = 0.527), while Ningqiang horse showed the lowest nucleotide diversity (${\pi}=0.00000$) and haplotype diversity (h = 0.000). The results also revealed that Chinese horses had a different polymorphic pattern of Y chromosome from European and American horses. In conclusion, Chinese horses revealed genetic diversity of Y chromosome, however more efforts should be made to better understand the domestication and paternal origin of Chinese indigenous horses.

Computational Analysis of Neighboring Genes on Arabidopsis thaliana Chromosomes 4 and 5: Their Genomic Association as Functional Subunits

  • Goh, Sung-Ho;Kim, Tae-Hyung;Kim, Jee-Hyub;Nam, DouGu;Choi, Doil;Hur, Cheol-Goo
    • Genomics & Informatics
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    • v.1 no.1
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    • pp.40-49
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    • 2003
  • The genes related to specific events or pathways in bacteria are frequently localized proximate to the genome of their neighbors, as with the structures known as operon, but eukaryotic genes seem to be independent of their neighbors, and are dispersed randomly throughout genomes. Although cases are rare, the findings from structures similar to prokaryotic operons in the nematode genome, and the clustering of housekeeping genes on human genome, lead us to assess the genomic association of genes as functional subunits. We evaluated the genomic association of neighboring genes on chromosomes 4 and 5 of Arabidopsis thaliana with and without respectively consideration of the scaffold/matrix­attached regions (S/MAR) loci. The observed number of functionally identical bigrams and trig rams were significantly higher than expected, and these results were verified statistically by calculating p-values for weighted random distributions. The observed frequency of functionally identical big rams and trig rams were much higher in chromosome 4 than in chromosome 5, but the frequencies with, and without, consideration of the S/MAR in each chromosome were similar. In this study, a genomic association among functionally related neighboring genes in Arabidopsis thaliana was suggested.

Genetic relationship of Aloe vera 'Saengjang', a new forma, based on cpDNA and ITS sequence variation (cpDNA와 ITS 염기변이에 근거한 신품종 생장알로에 유전적 상관관계)

  • Srikanth, Krishnamoorthy;Jang, Seon Il;Whang, Sung Soo
    • Korean Journal of Plant Taxonomy
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    • v.44 no.4
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    • pp.250-256
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    • 2014
  • This study was carried out to understand the genetic relationship of three Aloe spp. cultivated in Korea, A. saponaria, A. vera and A. arborescens and a new variant in Korea based on three plastid (matK, trnL-F, rbcL) and one nuclear (ITS regions) DNA barcode markers. A total of 2,420 bp sequence was amplified. Two indels were detected in the trnL region, and also several species specific nucleotide loci were detected in all 29 parsimonious informative sites, and 148 variable sites were detected among four taxa studied while 170 variable and 75 parsimonious sites were detected when other Aloe spp. in worldwide were used. An UPGMA phenogram with 10,000 bootstrap replication showed that the new variant was closest to A. vera. The variant was not morphologically and genetically concurrent with any reported species so far. The clustering of Aloe species were broadly in agreement with previously reported results.

Chromosome Imbalances and Alterations in the p53 Gene in Uterine Myomas from the Same Family Members: Familial Leiomyomatosis in Turkey

  • Hakverdi, Sibel;Demirhan, Osman;Tunc, Erdal;Inandiklioglu, Nihal;Uslu, Inayet Nur;Gungoren, Arif;Erdem, Duygu;Hakverdi, Ali Ulvi
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.651-658
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    • 2013
  • Uterine leiomyomas (UL) are extremely common neoplasms in women of reproductive age, and are associated with a variety of characteristic choromosomal aberrations (CAs). The p53 gene has been reported to play a crucial role in suppressing the growth of a variety of cancer cells. Therefore, the present study investigated the effects of CAs and the p53 gene on ULs. We performed cytogenetic analysis by G-banding in 10 cases undergoing myomectomy or hysterectomy. Fluorescence in situ hybridization (FISH) with a p53 gene probe was also used on interphase nuclei to screen for deletions. In patients, CAs were found in 23.4% of 500 cells analysed, significantly more frequent than in the control group (p<0.001). In the patients, 76% of the abnormalities were structural aberrations (deletions, translocations and breaks), and only 24% were numerical. Deletions were the most common structural aberration observed in CAs. Among these CAs, specific changes in five loci 1q11, 1q42, 2p23, 5q31 and Xp22 have been found in our patients and these changes were not reported previously in UL. The chromosome breaks were more frequent in cases, from high to low, 1, 2, 6, 9, 3, 5, 10 and 12. Chromosome 22, X, 3, 17 and 18 aneuploidy was observed to be the most frequent among all numerical aberrations. We observed a low frequency of p53 losses (2-11%) in our cases. The increased incidence of autosomal deletions, translocations, chromatid breaks and aneuploidy, could contribute to the progression of the disease along with other chromosomal alterations.

Pinus densiflora for. erecta - Can It Be Treated Genetically as a Distinct Group? - Reconsideration Based on Allozyme Data - (금강소나무 - 유전적으로 별개의 품종으로 인정될 수 있는가? - 동위효소분석 결과에 의한 고찰 -)

  • Kim, Zin Suh;Lee, Seok Woo;Hwang, Jae Woo;Kwon, Ki Won
    • Journal of Korean Society of Forest Science
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    • v.82 no.2
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    • pp.166-175
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    • 1993
  • The genetic variation patterns at 23 loci coding for 16 isozymes in eight natural populations of Pinus densiflora for. erects distributed in Kangwon-Kyungbuk region and 17 populations of Pinus densiflora and 13 populations of Pinus thunbergii were compared. The absence of marker alleles specific to P. thunbergii and almost the same allele-frequency distributions to those of P. densiflora did not support the hypothesis that P. densiflora for. erecta is a introgressive hybrid between P. densiflora and P. thunbergii. From the results of the hierarchial analysis of population differentiation using Wright's F statistics(1978), the frequency distributions of single-locus distance coefficients and other genetic analysis (genetic distance, cluster analysis, factor analysis, resin duct analysis), it was concluded that Pinus densiflora for. erecta cannot be treated genetically as a distinct group from other natural populations of P. densiflora.

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Reverse Random Amplified Microsatellite Polymorphism Reveals Enhanced Polymorphisms in the 3' End of Simple Sequence Repeats in the Pepper Genome

  • Min, Woong-Ki;Han, Jung-Heon;Kang, Won-Hee;Lee, Heung-Ryul;Kim, Byung-Dong
    • Molecules and Cells
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    • v.26 no.3
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    • pp.250-257
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    • 2008
  • Microsatellites or simple sequence repeats (SSR) are widely distributed in eukaryotic genomes and are informative genetic markers. Despite many advantages of SSR markers such as a high degree of allelic polymorphisms, co-dominant inheritance, multi-allelism, and genome-wide coverage in various plant species, they also have shortcomings such as low polymorphic rates between genetically close lines, especially in Capsicum annuum. We developed an alternative technique to SSR by normalizing and alternating anchored primers in random amplified microsatellite polymorphisms (RAMP). This technique, designated reverse random amplified microsatellite polymorphism (rRAMP), allows the detection of nucleotide variation in the 3' region flanking an SSR using normalized anchored and random primer combinations. The reproducibility and frequency of polymorphic loci in rRAMP was vigorously enhanced by translocation of the 5' anchor of repeat sequences to the 3' end position and selective use of moderate arbitrary primers. In our study, the PCR banding pattern of rRAMP was highly dependent on the frequency of repeat motifs and primer combinations with random primers. Linkage analysis showed that rRAMP markers were well scattered on an intra-specific pepper map. Based on these results, we suggest that this technique is useful for studying genetic diversity, molecular fingerprinting, and rapidly constructing molecular maps for diverse plant species.