• International Journal of Industrial Entomology and Biomaterials
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• 제25권1호
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• pp.81-92
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• 2012

A total of 85 Chinese-origin silkworm strains preserved in Korea were genotyped for eight polymorphic micro-satellite loci. We obtained per-locus number of alleles, ranging from 5 to 14 with an average value of 9.5, perlocus observed heterozygosity, ranging from 0.07 to 0.99, and per-locus polymorphic information content (PIC), ranging from 0.34 to 0.82, indicating that some loci are highly variable. Phylogenetic analysis with the eight concatenated microsatellite loci showed no clustering on the basis of known strain characteristics. A total of 22 strain-specific apomorphic alleles, which discriminate 19 among 85 silkworm strains were obtained from eight loci. These strain-specific alleles, thus, can casually be utilized for the discrimination of applicable strains without any further typing of other loci. Furthermore, a substantial number of homozygote strains, represented by 27 among 76 alleles in eight loci were found. These results collectively suggest that the silkworm microsatellite DNA is actually and potentially important molecular markers for the eventual discrimination of silkworm strains that are preserved as hundreds in Korea.

• 한국동물위생학회지
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• 제41권4호
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• pp.257-262
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• 2018

Avian pathogenic E. coli (APEC) causes severe economic losses in the poultry farms, due to systemic infections leading to lethal colisepticemia. It causes a variety of diseases from air sac infection to systemic spread leading to septicemia. Secondary infection contains opportunistic infections due to immunosuppression disease. Collibacillosis causes the great problems in the poultry industry in Korea. Thus, it is necessary to identify and classify the characteristics of E. coli isolate of chicken origin to confirm the diversity of symptoms and whether they are transmitted among the farms. Fragment analysis is identify the difference in the number of Variable-Number Tandem-Repeats (VNTRs) for genotyping. VNTRs have repeating structure (Microsatellite, Short tandem repeats; STR, Simple sequence repeats; SSR) in the chromosome. This region can be used as a genetic marker because of its high mutation rate. And various lengths of the amplified DNA fragment cause the difference in the number of repetition of the DNA specific site. The number of repetition sequences indicates the separated size of fragments, so the each fragments can be distinguished by specific samples. The results of the sample show that there is no difference in six microsatellite loci (yjiD, aidB, molR_1, ftsZ, b1668, yibA). There are differences among the farms in relation of the number of repetitions of other six microsatellite loci (ycgW, yaiN, yiaB, mhpR, b0829, caiF). Four (ycgW, yiaB, b0829, caiF) of these six microsatellite loci show statistically significant differences (P<0.05). It means that the analysis using four microsatellite loci including ycgW, yiaB, b0829, and caiF can confirm among the farms. Five E. coli samples in one farm have same SSR repetition at all markers. But, there are significant differences from other farms at Four (ycgW, yiaB, b0829, caiF) microsatellite loci. These results emphasize again that the four microsatellite loci makes a difference in the amplified DNA fragments, enabling it to be used for E. coli genotyping.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2015년도 춘계학술대회 및 임시총회
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• pp.42-42
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• 2015

Plasmodiophora brassicae, the causal agent of clubroot disease, does the most serious damage to the Brassica crops. The limited control approaches make that the identification of clubroot resistance (CR) is more important for developing CR cultivars of the Brassica crops. So far, 8 CR loci were mapped. However, the variation of P. brassicae leads to the rapid erosion of its resistance. To identify novel CR genes, we employed three mapping population, derived from crosses between Chinese cabbage and turnip inbred lines ($59-1{\times}ECD04$ and $BJN3-1{\times}Siloga$) or between Chinese cabbage inbred lines ($BJN3-1{\times}85-I-II$), to perform QTL analysis. Totally, 8 CR loci were indentified and showed race-specific resistance. Physical mapping of these 8 loci suggested that 4 were located previously mapped position, indicating they might be the same allele or different alleles of the same genes. Other 4 loci were found to be novel. Further, CR near isogenic line carrying each CR locus was developed based on the marker assisted selection. Verification of these CR loci was underway. Identification of these novel CR genes would facilitate to breed broad-spectrum and durable CR cultivars of B. rapa by pyramiding strategies.

• Asian-Australasian Journal of Animal Sciences
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• 제20권5호
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• pp.633-637
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• 2007

Genetic variation in Nilagiri sheep, the only apparel wool breed in South India was studied using 25 FAO recommended ovine-specific microsatellite markers. The number of observed alleles ranged from 3 to 8 with a mean of 5 across all loci. The size of alleles ranged from 72 to 228 bp. The frequency of alleles ranged from 0.0104 to 0.5781. In total, 125 alleles were observed at the 25 loci studied. The effective number of alleles ranged from 2.18 to 6.49. The mean number of effective alleles was 3.84 across all loci. All the 25 loci were found to be highly polymorphic. The PIC values ranged from 0.4587 to 0.8277 with a mean of 0.6485. Of 25 microsatellites studied, 17 were in Hardy-Weinberg Equilibrium proportions. The observed heterozygosity ranged from 0.4222 to 1.000 with a mean value of 0.7610 whereas the expected heterozygosity ranged from 0.5415 to 0.8459 with a mean value of 0.7213. Except six loci, the other loci revealed negative within-population inbreeding estimates (FIS) indicating excess of heterozygotes in the population of Nilagiri sheep.

• Plant Resources
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• 제1권2호
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• pp.81-91
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• 1998

Fifty-nine Korean soybean (Glycine max L. Merr.) landrace accessions were tested for genotype fingerprinting, differentiation and association between morphological traits and SSR profile. Using 8 SSR loci, 59 varieties were divided into 55 groups, and only 4 pairs of varieties were not uniquely identified. The resolving power of SSR for soybean genotyping was much higher than that of the morphological traits that were studied. Identification efficiency also differed among SSR loci. Those loci with higher numbers of alleles distinguished varieties more effectively. Genetic differentiation values of the soybean landraces varied from 0.57 to 0.82 with a mean of 0.68. The number of alleles detected by the 8 loci ranged from 3 to 8. and the effective number of alleles ranged from 2.3 to 5.1. In a study of the association of SSR alleles with morphological traits, some alleles seemed to be related with some specific morphological traits. Comparison of two kinds of dendrograms which were derived from SSR markers and quantitative traits indicated that the dendrograms were not consistent. Considering the correlation between single SSR locus and qualitative traits governed by major genes, the data suggest that alleles of microsatellite loci be more closely related to some traits determined by major genes than those determined by minor genes.

• Journal of Animal Science and Technology
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• 제48권5호
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• pp.637-650
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• 2006

한국의 예산과 당진에서 각각 채취된 붕어 (Carassius auratus)와 떡붕어 (Carassius cuvieri)로부터 genomic DNA를 분리 추출하여 반복해서 PCR로 증폭시켰다. 선택된 7개의 RAPD primer를 이용하여 primer 당 total loci, shared loci by each species, polymorphic 및 specific loci를 얻어냈다. 2종의 붕어로부터 primer와 2지역간에 banding patterns의 복잡성이 두드러지게 나타났다. DNA fragment의 분자적 크기는 150bp에서부터 1,600bp까지 커다란 차이를 나타내었다. 본 연구에서 CCY 붕어 종에서는 458개의 loci가 나타났고, CCD 떡붕어 종에서는 358개의 loci가 확인되었다. 또한 CCY 붕어 종에서는 84개의 polymorphic loci (18.3%)가 확인되었고, CCD떡붕어 종에서는 48개의 polymorphic loci (13.4%)가 확인되었다. CCY 붕어 종에서는 154개의 shared loci가 나타났으며, 이는 primer당 평균적으로 22개의 loci로 확인되었다. 또한 CCD떡붕어 종에서는 187개의 shared loci가 확인되었고, 평균해서 primer 당 26.7개의 loci가 나타났다. CCY붕어 종과 CCD 떡붕어 종의 polymorphic loci는 각각 84개와 48개로 확인되었다. 모든 붕어와 떡붕어 시료의 평균적인 BS value를 기초로 해서 CCY 붕어 종의 similarity matrix를 조사해 본 결과 0.434로부터 0.868까지 나타났고, CCD 떡붕어 종의 값은 0.449로부터 0.924까지 확인되었다. CCY 붕어 종내의 평균적인 BS value는 0.641±0.013이고, CCD 떡붕어 종내의 BS value의 평균값은 0.684±0.013을 나타내었다. 결과적으로 CCD 떡붕어 종내의 개체의 BS value 평균값이 CCY 붕어 종내의 평균값보다 높게 나타났다. 2 붕어와 떡붕어간의 평균적인 BS value은 0.484±0.007 (0.307～0.682)를 나타내었다. 7개의 primer를 사용하여 얻어진 dendrogram은 cluster 1 (AURATUS no. 01～AURATUS no. 11), cluster 2 (CUVIERI no. 12～CUVIERI no. 21) 및cluster 3 (CUVIERI no. 22)와 같이 3개의 유전적 클러스터로 나뉘어졌다. CCY 붕어 종내의 8번째 개체 (AURATUS no. 08)와 9번째 개체 (AURATUS no. 09) 사이가 가장 가까운 유전적 관계 (0.064)를 나타내었다. 또한 CCY붕어 종의 11번째(AURATUS no. 11)와 CCD떡붕어 종의 17번째 (CUVIERI no. 17) 사이가 가장 먼 유전적 거리 (0.477)를 나타내었다. 결과적으로 볼 때 한국 및 대서양산 lobster (0.612), 갈치 (0.708), 동자개(0.714)에 비해서 상대적으로 낮은 유전적 거리를 나타내었다.

• Parasites, Hosts and Diseases
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• 제38권1호
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• pp.45-48
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• 2000

An enzyme analysis of the liver fluke, Clonorchis sinensis from Kimhae, Korea and from Shenyang, China was conducted using a horizontal. starch gel electrophoresis in order to elucidate their genetic relationships. A total of eight enzymes was employed from two different kinds of buffer systems. Two loci from each enzyme of aconitase and esterase (${\alpha}-Na{\;}and{\;}{\beta}-Na$) : and only one locus each from six enzymes, gluucose-6-phosphate dehydrogenase (G6PD), ${\alpha}-glycerophosphate$ dehydrogenase (GPD), 3-hydroxybutyrate dehydrogenase (HBDH), malate dehydrogenase (MDH), phosphoglucose isomerase (PGI), and phosphoglucomutase (PGM) were detected. Most of loci in two populations of C. sinensis showed homozygous monomorphic banding patterns and one of them, GPD was specific as genetic markers between two different populations. However, esterase (${\alpha}-Na$), GPD, HBDH and PGI loci showed polymorphic banding patterns. Two populations of C. sinensis were more closely clustered within the range of genetic identity value of 0.998-1.0. In summarizing the above results, two populations of C. sinensis employed in this study showed mostly monomorphic enzyme protein banding patterns, and genetic differences specific between two populations.

• 한국발생생물학회지:발생과생식
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• 제10권4호
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• pp.227-238
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• 2006

한반도의 동쪽에 위치해 있는 삼척(venus clam from Samcheok; VCS)과 원산(venus clam from Wonsan; VCW) 지역에서 채취된 민들조개(Gomphina aequilatera)에서 genomic DNAs(gDNAs)를 분리 추출하였다. 증폭산물은 primer agarose 전기영동법에 의해서 생성되었고, EtBr에 의해서 염색된 이후에 자외선에 의해서 확인되었다. 150 bp에서 2,400 bp에 해당되는 shared loci, polymorphic 및 specific loci를 얻기 위해서 BION-21, BION-23, BION-25, BION-27, BION-29, BION-31 및 BION-33와 같은 7개의 primer를 사용하였다. 본 연구에서 7개의 primer는 VCS 민들조개 집단에서 147개의 polymorphic loci(147/954 loci, 15.41%)와 VCW 집단에서 274개의 polymorphic loci(274/996 loci, 27.51%)를 확인하였다. 이것은 VCS 민들조개 집단에서 보다 VCW 집단에서 더 높은 유전적 변이를 나타내고 있다는 것을 제시하고 있다. 특히 BION-21 primer에 의해서 나타난 700 bp는 민들조개 2개 집단에서 공통적으로 확인되었으며, 이러한 것은 집단이나 종을 확인할 수 있는 marker로서 활용이 가능할 것이다. 이러한 특이한 primer는 개체, 종 및 집단에서 서로 다른 DNA 다형성을 나타내며, 개체나 집단을 확인하는 데 유용하다는 것을 알 수 있다. 2개 민들조개 집단의 개체들을 비교해 보았을 때 SAMCHEOK no. 03와 WONSAN no. 22에서 가장 긴 유전적 거리(0.696)를 나타내었다. 3개의 genetic groupings and dendrogram을 포함한 complete linkage cluster analysis을 통해서 볼 때 지리적 거리가 있었지만 삼척과 원산 2 민들조개 집단의 개체 정체성과 다소 가까운 친척관계를 확인시켜 주었다. 분자적인 표지인자로부터 얻어진 종내 분류와 clustering analyses은 패각 크기, 패각 형태 및 패각 색깔과 같은 형태적인 형질을 기초한 재래적인 종 분류를 지원하고 있다. 따라서 위에서 언급된 바와 같이 RAPD 분석은 VCS 민들조개 집단이 VCW 집단과 어느 정도 차이가 있다는 것을 확인시켜 주었다.

• Molecules and Cells
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• 제44권9호
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• pp.627-636
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• 2021

The three-dimensional organization of chromatin and its time-dependent changes greatly affect virtually every cellular function, especially DNA replication, genome maintenance, transcription regulation, and cell differentiation. Sequencing-based techniques such as ChIP-seq, ATAC-seq, and Hi-C provide abundant information on how genomic elements are coupled with regulatory proteins and functionally organized into hierarchical domains through their interactions. However, visualizing the time-dependent changes of such organization in individual cells remains challenging. Recent developments of CRISPR systems for site-specific fluorescent labeling of genomic loci have provided promising strategies for visualizing chromatin dynamics in live cells. However, there are several limiting factors, including background signals, off-target binding of CRISPR, and rapid photobleaching of the fluorophores, requiring a large number of target-bound CRISPR complexes to reliably distinguish the target-specific foci from the background. Various modifications have been engineered into the CRISPR system to enhance the signal-to-background ratio and signal longevity to detect target foci more reliably and efficiently, and to reduce the required target size. In this review, we comprehensively compare the performances of recently developed CRISPR designs for improved visualization of genomic loci in terms of the reliability of target detection, the ability to detect small repeat loci, and the allowed time of live tracking. Longer observation of genomic loci allows the detailed identification of the dynamic characteristics of chromatin. The diffusion properties of chromatin found in recent studies are reviewed, which provide suggestions for the underlying biological processes.

• BMB Reports
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• 제28권4호
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• pp.359-362
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• 1995

The highly polymorphic variable number of tandem repeat (VNTR) loci in the human genome are informative markers for the genetic characterization of individuals in the paternity test and forensic science as well as for the study of human disease. In this study, VNTR loci D1S80 and D2S123 have been amplified by PCR and the amplified length polymorphic alleles were detected with a discontinuous vertical PAGE system and silver staining. For explicit DNA typing, PCR optimization, in which amplification efficiencies are similar over a wide range of allele sizes, non-specific amplifications are minimal, and new longer alleles have high amplification efficiency, has been performed by changing the PCR reaction buffer composition and thermal cycling conditions. It turned out that adding an appropriate amount of Tween 20 and NP40 to the PCR reaction buffer and raising the annealing temperature to $68^{\circ}C$ in thermal cycling made it possible for optimal VNTR loci amplification. A modified PAGE system for VNTR separation was established. Under these conditions, new longer alleles in the 01580 locus were discovered and 025123 pattern changes in colorectal tumors were observed. These technical tips are valuable for detecting various amplified fragment length polymorphisms.