• Title/Summary/Keyword: Microsatellite DNA

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Colorectal Carcinoma in Malaysians: DNA Mismatch Repair Pattern in a Multiethnic Population

  • Cheah, Phaik-Leng;Looi, Lai-Meng;Teoh, Kean-Hooi;Rahman, Nazarina Abdul;Wong, Li-Xuan;Tan, Soo-Yong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.7
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    • pp.3287-3291
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    • 2014
  • Background: The interesting preponderance of Chinese with colorectal carcinoma (CRC) amongst the three major ethnic groups in Malaysia prompted a study to determine DNA mismatch repair (MMR) status in our CRC and attempt correlation with patient age, gender and ethnicity as well as location, grade, histological type and stage of tumour. Histologically re-confirmed CRC, diagnosed between $1^{st} $January 2005 and $31^{st}$ December 2007 at the Department of Pathology, University of Malaya Medical Centre, were immunohistochemically stained with monoclonal antibodies to MMR proteins, MLH1, MSH2, MSH6 and PMS2 on the Ventana Benchmark XT autostainer. Of the 142 CRC cases entered into the study, there were 82 males and 60 females (M:F=1.4:1). Ethnically, 81 (57.0%) were Chinese, 32 (22.5%) Malays and 29 (20.4%) Indians. The patient ages ranged between 15-87 years (mean=62.4 years) with 21 cases <50-years and 121 ${\geq}50$-years of age. 14 (9.9%) CRC showed deficient MMR (dMMR). Concurrent loss of MLH1 and PMS2 occurred in 10, MSH2 and MSH6 in 2 with isolated loss of MSH6 in 1 and PMS2 in 1. dMMR was noted less frequently amongst the Chinese (6.2%) in comparison with their combined Malay and Indian counterparts (14.8%), and was associated with right sided and poorly differentiated tumours (p<0.05). 3 of the 5 (60.0%) dMMR CRC cases amongst the Chinese and 1 of 9 cases (11.1%) amongst the combined Malay and Indian group were <50-years of age. No significant association of dMMR was noted with patient age and gender, tumour stage or mucinous type.

Genetic Traceability of Black Pig Meats Using Microsatellite Markers

  • Oh, Jae-Don;Song, Ki-Duk;Seo, Joo-Hee;Kim, Duk-Kyung;Kim, Sung-Hoon;Seo, Kang-Seok;Lim, Hyun-Tae;Lee, Jae-Bong;Park, Hwa-Chun;Ryu, Youn-Chul;Kang, Min-Soo;Cho, Seoae;Kim, Eui-Soo;Choe, Ho-Sung;Kong, Hong-Sik;Lee, Hak-Kyo
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.7
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    • pp.926-931
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    • 2014
  • Pork from Jeju black pig (population J) and Berkshire (population B) has a unique market share in Korea because of their high meat quality. Due to the high demand of this pork, traceability of the pork to its origin is becoming an important part of the consumer demand. To examine the feasibility of such a system, we aim to provide basic genetic information of the two black pig populations and assess the possibility of genetically distinguishing between the two breeds. Muscle samples were collected from slaughter houses in Jeju Island and Namwon, Chonbuk province, Korea, for populations J and B, respectively. In total 800 Jeju black pigs and 351 Berkshires were genotyped at thirteen microsatellite (MS) markers. Analyses on the genetic diversity of the two populations were carried out in the programs MS toolkit and FSTAT. The population structure of the two breeds was determined by a Bayesian clustering method implemented in structure and by a phylogenetic analysis in Phylip. Population J exhibited higher mean number of alleles, expected heterozygosity and observed heterozygosity value, and polymorphism information content, compared to population B. The $F_{IS}$ values of population J and population B were 0.03 and -0.005, respectively, indicating that little or no inbreeding has occurred. In addition, genetic structure analysis revealed the possibility of gene flow from population B to population J. The expected probability of identify value of the 13 MS markers was $9.87{\times}10^{-14}$ in population J, $3.17{\times}10^{-9}$ in population B, and $1.03{\times}10^{-12}$ in the two populations. The results of this study are useful in distinguishing between the two black pig breeds and can be used as a foundation for further development of DNA markers.

Genetic Contribution of Indigenous Yakutian Cattle to Two Hybrid Populations, Revealed by Microsatellite Variation

  • Li, M.H.;Nogovitsina, E.;Ivanova, Z.;Erhardt, G.;Vilkki, J.;Popov, R.;Ammosov, I.;Kiselyova, T.;Kantanen, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.613-619
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    • 2005
  • Indigenous Yakutian cattle' adaptation to the hardest subarctic conditions makes them a valuable genetic resource for cattle breeding in the Siberian area. Since early last century, crossbreeding between native Yakutian cattle and imported Simmental and Kholmogory breeds has been widely adopted. In this study, variations at 22 polymorphic microsatellite loci in 5 populations of Yakutian, Kholmogory, Simmental, Yakutian-Kholmogory and Yakutian-Simmental cattle were analysed to estimate the genetic contribution of Yakutian cattle to the two hybrid populations. Three statistical approaches were used: the weighted least-squares (WLS) method which considers all allele frequencies; a recently developed implementation of a Markov chain Monte Carlo (MCMC) method called likelihood-based estimation of admixture (LEA); and a model-based Bayesian admixture analysis method (STRUCTURE). At population-level admixture analyses, the estimate based on the LEA was consistent with that obtained by the WLS method. Both methods showed that the genetic contribution of the indigenous Yakutian cattle in Yakutian-Kholmogory was small (9.6% by the LEA and 14.2% by the WLS method). In the Yakutian-Simmental population, the genetic contribution of the indigenous Yakutian cattle was considerably higher (62.8% by the LEA and 56.9% by the WLS method). Individual-level admixture analyses using STRUCTURE proved to be more informative than the multidimensional scaling analysis (MDSA) based on individual-based genetic distances. Of the 9 Yakutian-Simmental animals studied, 8 showed admixed origin, whereas of the 14 studied Yakutian-Kholmogory animals only 2 showed Yakutian ancestry (>5%). The mean posterior distributions of individual admixture coefficient (q) varied greatly among the samples in both hybrid populations. This study revealed a minor existing contribution of the Yakutian cattle in the Yakutian-Kholmogory hybrid population, but in the Yakutian-Simmental hybrid population, a major genetic contribution of the Yakutian cattle was seen. The results reflect the different crossbreeding patterns used in the development of the two hybrid populations. Additionally, molecular evidence for differences among individual admixture proportions was seen in both hybrid populations, resulting from the stochastic process in crossing over generations.

Possibility of Natural Hybridization between Red Seabream (Pagrus major) and Blackhead Seabream (Acanthopagrus schlegeli) (참돔(Pagrus major)과 감성돔(Acanthopagrus schlegeli) 종간 자연 잡종 가능성)

  • Kang, Jung-Ha;Yang, Sang-Geun;Kim, Eun-Mi;Noh, Eun-Soo;Kim, Dong-Gyun;Kim, Bong-Seok;Choi, Tae-Jin
    • Journal of Life Science
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    • v.25 no.1
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    • pp.16-20
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    • 2015
  • During the storage of these two species in a large conservation tank, fertilized eggs were collected and the offspring were raised. During culturing of the offspring, individuals with mixed characteristics of these two species were observed, and 96 individuals were randomly tested using microsatellite markers applicable to both species. Among the 96 individuals, 15 individuals with mixed morphological characteristics were confirmed to be hybrids showing both of genotypes red seabream and blackhead seabream. Additionally, based on sequence analysis of mitochondrial cytochrome oxidase subunit 1 (mtDNA CO1), 81 showed 99% nucleotide sequence identity to that of black sea bream, and the remaining 15 individuals showed over 99% sequence identity to that of red seabream. So, hybrids were produced by female red seabream and male blackhead seabream. These results suggest that hybrids may form in nature between these two species if their habitats overlap due to the influence of humans or global climate change.

Application of DNA Test for Individual Traceability in Hanwoo (Korean Cattle) (한우의 개체 추적 검증을 위한 유전자 감식 기법 활용 연구)

  • 이학교;전광주;공홍식;오재돈;최일신;김종대;조창연;윤두학;신형두
    • Food Science of Animal Resources
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    • v.24 no.1
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    • pp.8-14
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    • 2004
  • Identification of animals has been made with an ear tag with dummy code, and blood typing has been used for paternity and individual identification in live animals. As various genetic markers are for different cattle breeds vary, the discrete genetic markers are necessary to identify Hanwoo. A total of 740 progeny testing Hanwoo were used to identify Hanwoo specific markers. To examine traceability of individuals by using breed specific genetic codes, four animal were randomly sampled, and traced from live animals to post-slaughter processing stages. The candidate genetic makers used in the study were 16 DNA microsatellites which were identified in romosomes 1 and 14. The number of alleles of those DNA microsatellites ranged from a minimum of 3 to maximum of 12. The heterozygote frequency ranged from 0.022 to 0.824. Effective number of alleles for each DNA microsatellites were 3 to 6. Six selected candidate genetic markers were able ti trace individual cattle with an 100% confidence level.

Construction of Genetic Linkage Map and Identification of Quantitative Trait Loci in Populus davidiana using Genotyping-by-sequencing (Genotyping-by-sequencing 기법을 이용한 사시나무(Populus davidiana) 유전연관지도 작성 및 양적형질 유전자좌 탐색)

  • Suvi Kim;Yang-gil Kim;Dayoung Lee;Hye-jin Lee;Kyu-Suk Kang
    • Journal of Korean Society of Forest Science
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    • v.112 no.1
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    • pp.40-56
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    • 2023
  • Tree species within the Populus genus grow rapidly and have an excellent capacity to absorb carbon, conferring substantial ability to effective purify the environment. Poplar breeding can be achieved rapidly and efficiently if a genetic linkage map is constructed and quantitative trait loci (QTLs) are identified. Here, a high-density genetic linkage map was constructed for the control pollinated progeny using the genotyping-by-sequencing (GBS) technique, which is a next-generation sequencing method. A search was also performed for the genes associated with quantitative traits located in the genetic linkage map by examining the variables of height and diameter at root collar, and resilience to insect damage. The height and diameter at root collar were measured directly, while the ability to recover from insect damage was scored in a 4-year-old breeding population of aspen hybrids (Odae19 × Bonghyeon4 F1) established in the research forest of Seoul National University. After DNA extraction, paternity was confirmed using five microsatellite markers, and only the individuals for which paternity was confirmed were used for the analysis. The DNA was cut using restriction enzymes and the obtained DNA fragments were prepared using a GBS library and sequenced. The analyzed results were sorted using Populus trichocarpa as a reference genome. Overall, 58,040 aligned single-nucleotide polymorphism (SNP) markers were identified, 17,755 of which were used for mapping genetic linkages. The genetic linkage map was divided into 19 linkage groups, with a total length of 2,129.54 cM. The analysis failed to identify any growth-related QTLs, but a gene assumed to be related to recovery from insect damage was identified on linkage group (chromosome) 4 through genome-wide association study.

Genetic control of shell color variation in the Haliotis discus hannai by mating experiments (교배실험을 통한 북방전복 (Haliotis discus hannai)의 패각색 변이에 대한 유전적 지배)

  • Park, Choul Ji;Nam, Won Shik;Lee, Myeong Seok;Kang, Ji-Yun;Kim, Kyung Kil
    • The Korean Journal of Malacology
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    • v.30 no.4
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    • pp.409-413
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    • 2014
  • Purple-colored shell individuals were discovered among normal green-colored shell individuals in artificial seed of Pacific abalone, Haliotis discus hannai, reared on an ordinary type of diatom and artificial diet. In the present study, factorial mating experiments were designed to clarify the genetic control of the variant (purple type) and normal (green type) of shell color. The parental population of purple type and green type individuals were derived from a single family between a female and male of each type of coloration. The all mating families were reared in same tank for the same breed environment. The individual of 4 type families were distinguished by paternity test using microsatellite DNA. In factorial mating experiments, all individuals offspring of GG (green type female and green type male), GP (green type female and purple type male) and PG (purple type female and green type male) mating types appeared to green type. In only PP (purple type female and purple type male) mating type, all individuals offspring appeared to purple type. The results suggested that the purple shell color is controlled by recessive purple type allele and a dominant green type allele at a single locus.

Evaluation of MT1XT20 Single Quasi-Monomorphic Mononucleotide Marker for Characterizing Microsatellite Instability in Persian Lynch Syndrome Patients

  • Farahani, Najmeh;Nikpour, Parvaneh;Emami, Mohammad Hassan;Hashemzadeh, Morteza;Zeinalian, Mehrdad;Shariatpanahi, Seyed Shervin;Salehi, Rasoul
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.9
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    • pp.4259-4265
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    • 2016
  • Background: Colorectal malignancies with high microsatellite instability (MSI-H), either hereditary (Lynch syndrome) or sporadic, demonstrate better prognosis and altered response to 5FU chemotherapy. It is now recommended to perform MSI testing for all new cases of colorectal cancer regardless of being categorized as hereditary or sporadic. For MSI detection, immunohistochemistry or PCR-based protocols using a cohort of various sets of STR markers are recommended. Here we aimed to evaluate a simplified protocol using just a single STR marker, MT1XT20 mononucleotide repeat, for detection of MSI in Lynch syndrome patients. A Promega five-marker MSI testing panel and immunohistochemistry (IHC) were used as the gold standard in conjunction with MT1XT20. Materials and Methods: Colorectal patients with a positive history of familial cancers were selected by evaluating medical records. Based on Amsterdam II criteria for Lynch syndrome 20 families were short listed. DNA was extracted from formalin fixed paraffin embedded tumour and adjacent normal tissues resected from the index case in each family. Extracted DNA was subjected to MT1XT20 mononucleotide marker analysis and assessment with a commercially available five marker MSI testing kit (Promega, USA). IHC also was performed on tissue sections and the results were compared with PCR based data. Results: Eight (40%), seven (35%) and five (25%) cases were MSI positive using with the Promega kit, IHC and MT1XT20, respectively. Among the markers included in Promega kit, BAT26 marker showed instability in all 8 samples. NR24 and NR21 markers showed instability in 7 (87.5%), and BAT25 and MONO 27 in 6 (75%) and 5 (62.5%). Conclusions: Although MT1XT20 was earlier reported as a valid standalone marker for MSI testing in CRC patients, we could not verify this in our Iranian patients. Instead BAT26 among the markers included in Promega MSI testing kit showed instability in all 8 MSI-H CRC samples. Therefore, it seems BAT26 could act well as a single marker for MSI testing in Iranian CRC patients.

Genetic Relationship between Populations and Analysis of Genetic Structure in the Korean Native Chicken and the Endemic Chicken Breeds (한국재래닭 및 토착화 품종간의 유연 관계 및 유전 특성 분석)

  • Oh, J.D.;Kang, B.S.;Kim, H.K.;Park, M.N.;Chae, E.J.;Seo, O.S.;Lee, H.K.;Jeon, G.J.;Kong, H.S.
    • Korean Journal of Poultry Science
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    • v.35 no.4
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    • pp.361-366
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    • 2009
  • The purpose of this study was to assess the genetic variation and establish the relationship amongst breeds and strains using 7 chicken specific microsatellite markers. A total of 317 DNA samples from four Korean native chicken (KNC) strains (KR: Korean Native Red chicken strain, KY: Korean Native Yellow chicken strain, KL: Korean Native Black chicken strain, KO: Ogol chicken strain) and three introduced endemic chicken breeds (LE: Leghorn chicken breed, RI: Rhode Island Red chicken breed, CO: Cornish chicken breed). The size of microsatellite markers was decided using GeneMapper Software (v.4.0) after being analyzed using an ABI 3130 Genetic Analyzer. Frequencies of microsatellites markers were used to estimate heterozygosities and genetic distances. The lowest distance (0.074) was observed between the KY and KL breeds and the highest distance (0.779) between the KL and LE breeds. The KNC strains (KR, KY, KL) have comparatively near genetic distance each other. On the other side, each individual was not ramified to different groups and were spread evenly in phylogenetic dendrogram about all the KNC of each strain populations. But the endemic breed populations (LE, RI, CO) were ramified to different groups. The microsatellite polymorphism data were shown to be useful for assessing the genetic relationship between Korean native strains and other foreign breeds.

Expansion of an invasive species, Ailanthus altissima, at a regional scale in Japan

  • Chuman, Misaki;Kurokochi, Hiroyuki;Saito, Yoko;Ide, Yuji
    • Journal of Ecology and Environment
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    • v.38 no.1
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    • pp.47-56
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    • 2015
  • Ailanthus altissima, which is recognized as an invasive tree in the Western world, has been widely observed in Japan. To investigate how A. altissima expanded within-population and to new populations within a region, 446 A. altissima trees were sampled from three separate sites (A, B, and C) including 35 distantly positioned patches, with three chloroplast DNA markers and nine nuclear microsatellite markers. We detected 2, 2, and 3 chloroplast haplotypes in sites A, B, and C, respectively. In addition, 271, 40, and 41 nuclear genotypes were detected in sites A, B, and C, respectively. The clonal richness value was 0.85, 0.78, and 0.53 in sites A, B, and C, respectively. Most trees with the same genotypes were distributed in the same patch, indicating that range expansion by asexual reproduction was limited to a maximum of 45 meters. According to autocorrelation analysis, the extent of nonrandom spatial genetic structure was approximately 0-2 km in sites A and C. KINGROUP analyses showed that 812, 74, and 111 nuclear genotype pairs were detected to have kinship in sites A, B, and C, respectively. Most nuclear genotype pairs were detected within the same patches or sites. These results indicate that the number of A. altissima trees gradually increased from seeds, some of which were produced by trees within sites, meaning that this species could regenerate naturally. This shows the need for the future management of A. altissima as an invasive species in Japan.