• Title/Summary/Keyword: Marker Loci

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Genotyping of avian pathogenic Escherichia coli by DNA fragment analysis for the differences in simple sequence repeats

  • Han, Mi Na;Byeon, Hyeon Seop;Han, Seong Tae;Jang, Rae Hoon;Kim, Chang Seop;Choi, Seok Hwa
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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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.

Genetic Variability Based on Randomly Amplified Polymorphic DNA in Kacip Fatimah (Labisia pumila Benth & Hook f) collected from Melaka and Negeri Sembilan States of Malaysia

  • Bhore, Subhash J.;Nurul, A.H.;Shah, Farida H.
    • Journal of Forest and Environmental Science
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    • v.25 no.2
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    • pp.93-100
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    • 2009
  • In Malaysia, Labisia pumila Benth & Hook f, popularly known as 'Kacip Fatimah' has been used traditionally to treat various elements of the woman's health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-$40^{\circ}C$ were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.

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Isozyme Analysis on the Allotriploid between Rainbow Trout (Oncorhynchus mykiss) and Coho Salmon (O. kisutch) (무지개송어와 은연어간 잡종3배체의 부화자어에 대한 동위효소 분석)

  • HONG Kyung Pyo;MYOUNG Jung-Goo;KIM Pyong Kih;SON Jin-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.2
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    • pp.256-261
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    • 1996
  • For the purpose of identification of inheritance in allotriploid between rainbow trout (Oncorhynchus mykiss) and coho salmon (O. kisutch), five isozymes, lactate dehydrogenase (LDH), malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH), phosphoglucose isomerase (PGI) and phosphoglucomutase (PGM) from skeletal muscle in two species and their allotriploid were analyzed. All of these loci showed differences between two species and their allotriploid except PGI. Generally, coho salmon was more monomorphic in these isozyme loci than rainbow trout. Their allotriploids showed intermediate patterns between the parental species in those isozyme loci except PGI. As a result of this study, LDH, MDH, IDH and PSM may be used as useful genetic markers in these two species, and they also be of use in studying hybrid and allotriploid in salmonids.

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Detection of Quantitative Trait Loci Affecting Fat Deposition Traits in Pigs

  • Choi, B.H.;Lee, K.T.;Lee, H.J.;Jang, G.W.;Lee, H.Y.;Cho, B.W.;Han, J.Y.;Kim, T.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.11
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    • pp.1507-1510
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    • 2012
  • Quantitative trait loci (QTL) associated with fat deposition traits in pigs are important gene positions in a chromosome that influence meat quality of pork. For QTL study, a three generation resource population was constructed from a cross between Korean native boars and Landrace sows. A total of 240 F2 animals from intercross of F1 were produced. 80 microsatellite markers covering chromosomes 1 to 10 were selected to genotype the resource population. Intervals between adjacent markers were approximately 19 cM. Linkage analysis was performed using CRIMAP software version 2.4 with a FIXED option to obtain the map distances. For QTL analysis, the public web-based software, QTL express (http://www.qtl.cap.ed.ac.uk) was used. Two significant and two suggestive QTL were identified on SSC 6, 7, and 8 as affecting body fat and IMF traits. For QTL affecting IMF, the most significant association was detected between marker sw71 and sw1881 on SSC 6, and a suggestive QTL was identified between sw268 and sw205 on SSC8. These QTL accounted for 26.58% and 12.31% of the phenotypic variance, respectively. A significant QTL affecting IMF was detected at position 105 cM between markers sw71 and sw1881 on SSC 6.

Loss of Heterozygosity (LOH) on 17th and 18th Chromosome from Colorectal Carcinoma (대장암에서 17, 18번 염색체의 이형접합성 소실)

  • Lee, Jae-Sik
    • Korean Journal of Clinical Laboratory Science
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    • v.40 no.1
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    • pp.41-47
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    • 2008
  • Colorectal carcinoma is occurred frequently to Korean and so ranked the fourth from various cancers. Due to western dietary life, this cancer has been increased continually. Therefore, the study will be needed to find a candidate gene involved in the development and progression of colorectal carcinoma and to diagnose and treatment helpfully. The striking feature from cancer suppressor genes is known for LOH (loss of heterozygosity), which is the method to find allele genetic loss or mutation of cancer cell. The purpose of this study was designed to find a carcinogenic gene from colon cancer using microsatellite marker on 17th and 18th chromosome from 30 subjects. The LOH was investigated in order of D18S59 57% (17/30), TP53CA 50% (15/30), D18S68 47% (14/30), D18S69 43% (13/30). The genetic mutation depends on loci of colorectal carcinoma was shown higher with 2.44 from colon cancer than with 1.25 from right colorectal carcinoma (p<0.032). The genetic mutation with lymph nodes was investigated higher with 2.69 at mutated group than with 1.14 at non-mutated group (p<0.003). At genetic mutated pattern depends on disease stage, there was higher significant difference at III-IV stage 2.50 than that of I-II stage 1.17, respectively (p=0.015). There was no difference at comparison between histological classification and serological CEA increase. The loss on 18q21 found in this study is highly recurrence loci and was observed 43% for Korean with high recurrence. Therefore, LOH is a very useful tool to detect 18q21 loci in clinical application, prior to the treatment of colorectal carcinoma. After the operation of colorectol carcinoma, the efficient application using LOH at operated part tissue which is designed to protect the recurrence as well as its cure will be needed.

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SLA Homozygous Korean Native Pigs and Their Inbreeding Status Deduced from the Microsatellite Marker Analysis

  • Jung, Woo-Young;Lim, Hyun-Tae;Lim, Jae-Sam;Kim, Sung-Bok;Jeon, Jin-Tae;Lee, Jun-Heon
    • Journal of Animal Science and Technology
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    • v.52 no.6
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    • pp.451-457
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    • 2010
  • The porcine MHC (Major Histocompatibility Complex), encoding the SLA (Swine Leukocyte Antigen) genes, is one of the most significant regions associated with immune rejection in relation to transplantation. In this study, three SLA class I (SLA-1, SLA-3, SLA-2) loci and three SLA class II (DRB1, DQB1, DQA) loci were investigated in the previously unidentified Korean native pig (KNP) population that was closely inbred in the Livestock Technology Research Station in Cheongyang, Korea. Total thirteen KNPs from four generations were genotyped for the SLA alleles and haplotypes were investigated using PCR-SSP (Sequence-Specific Primer) method. The results showed that all of these KNPs had Lr-56.30/56.30 homozygous haplotype, indicating high level of inbreeding in the SLA genes. The inbreeding status of these animals was also investigated using microsatellite (MS) markers. From the 50 MS markers investigated, 17 MS markers were fixed in all generations and the fixed alleles are increased as 26 loci for the fourth generation. Two MS markers, S0069 and SW173, were heterozygous for all the animals tested. Observed and expected heterozygosities were calculated and the average inbreeding coefficients for each generation were also calculated. In the fourth generation, the average inbreeding coefficients was 0.732 and this may increase with further inbreeding process. Analysis of the SLA haplotypes and MS alleles can give important information for breeding the pigs for xenotransplantation studies.

Morphometric variation, genetic diversity and allelic polymorphism of an underutilised species Thaumatococcus daniellii population in Southwestern Nigeria

  • Animasaun, David Adedayo;Afeez, Azeez;Adedibu, Peter Adeolu;Akande, Feyisayo Priscilla;Oyedeji, Stephen;Olorunmaiye, Kehinde Stephen
    • Journal of Plant Biotechnology
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    • v.47 no.4
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    • pp.298-308
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    • 2020
  • Genetic diversity among Thaumatococcus daniellii populations in the southwestern region of Nigeria were assessed using morphometric and molecular markers to determine the population structure and existing genetic relationship for its improvement, conservation and sustainable utilisation. Populations from five locations in each of the six states were used for the study. Morphometric data were collected on folia characters and analysed for variability. Genome DNA was isolated from the plant leaf and amplified by polymerase chain reaction with inter-simple sequence repeat markers (ISSR) to determine the allelic polymorphism, marker effectiveness and genetic relationship of the population. The results showed significant variations in petiole length and leaf dimensions of the populations within and across the states. These morphometric traits are the major parameters that delimit the populations and they correlated significantly at P≤0.05. Analysis of the electrophoregram showed that the ISSR markers are effective for the diversity study. A total of 136 loci were amplified with an average of 7.16 loci per marker, 63.2% of the loci were polymorphic. The Principal Coordinate Analysis revealed that seven factors accounted for 81.6% of the variation and the dendrogram separated the populations into two major groups at a genetic distance of 10 (about 90% similarity) with sub-groups and clusters. Most populations within the state had a high degree of similarity, nonetheless, strong genetic relationship exists among populations from different states. The close relationship between populations across the states suggests a common progenitor, which are likely separated by ecological or geographical isolation mechanisms.

Genetic Improvement of Maize by Marker-Assisted Breeding (분자마커를 활용한 옥수수 육종)

  • Kim, Jae Yoon;Moon, Jun-Cheol;Baek, Seong-Bum;Kwon, Young-Up;Song, Kitae;Lee, Byung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.2
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    • pp.109-127
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    • 2014
  • Maize is one of the most important food and feed crops in the world including Southeast Asia. In spite of numberous efforts with conventional breeding, the maize productions remain low and the loss of yields by drought and downy mildew are still severe in Asia. Genetic improvement of maize has been performed with molecular marker and genetic engineering. Because maize is one of the most widely studied crop for its own genome and has tremendous diversity and variant, maize is considered as a forefront crop in development and estimation of molecular markers for agricultural useful trait in genetics and breeding. Using QTL (Quantitative Trait Loci) and MAS (Marker Assisted Breeding), molecular breeders are able to accelerate the development of drought tolerance or downy mildew resistance maize genotype. The present paper overviews QTL/MAS approaches towards improvement of maize production against drought and downy mildew. We also discuss here the trends and importance of molecular marker and mapping population in maize breeding.

Development of Novel Microsatellite Markers for Strain-Specific Identification of Chlorella vulgaris

  • Jo, Beom-Ho;Lee, Chang Soo;Song, Hae-Ryong;Lee, Hyung-Gwan;Oh, Hee-Mock
    • Journal of Microbiology and Biotechnology
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    • v.24 no.9
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    • pp.1189-1195
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    • 2014
  • A strain-specific identification method is required to secure Chlorella strains with useful genetic traits, such as a fast growth rate or high lipid productivity, for application in biofuels, functional foods, and pharmaceuticals. Microsatellite markers based on simple sequence repeats can be a useful tool for this purpose. Therefore, this study developed five novel microsatellite markers (mChl-001, mChl-002, mChl-005, mChl-011, and mChl-012) using specific loci along the chloroplast genome of Chlorella vulgaris. The microsatellite markers were characterized based on their allelic diversities among nine strains of C. vulgaris with the same 18S rRNA sequence similarity. Each microsatellite marker exhibited 2~5 polymorphic allele types, and their combinations allowed discrimination between seven of the C. vulgaris strains. The two remaining strains were distinguished using one specific interspace region between the mChl-001 and mChl-005 loci, which was composed of about 27 single nucleotide polymorphisms, 13~15 specific sequence sites, and (T)n repeat sites. Thus, the polymorphic combination of the five microsatellite markers and one specific locus facilitated a clear distinction of C. vulgaris at the strain level, suggesting that the proposed microsatellite marker system can be useful for the accurate identification and classification of C. vulgaris.

Quantitative Trait Loci for Stem Length in Soybean Using a Microsatellite Markers (콩에서 Microsatellite 마커를 이용한 양적형질 유전자의 분석)

  • Kim, Hyeun-Kyeung;Kang, Sung-Taeg;Kong, Hyeun-Jong;Park, In-Soo
    • Journal of Life Science
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    • v.14 no.2
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    • pp.339-344
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    • 2004
  • Identification of individual quantitative trait loci (QTL) is a prerequisite to application of marker-assisted selection for stern length. Two simple sequence repeat (SSR)-based linkage maps were constructed from recombination inbred line populations between cross of Keunolkong and Shinpaldalkong. Two parents used differed greatly in stem length, which were 30.57 cm and 49.75 cm in Keunolkong and Shinpaldalkong, respectively. Using the constructed maps, regression analysis and interval mapping were performed to identify QTLs conferring stem length. Four QTLs for stem length on linkage groups (LG) F, J, N and O were identified in the Keunolkong ${\times}$ Shinpaldalkong population and they totally explained 37.83% of variation for stem length. In the population, two major QTLs on LG J and O conditioning 14.25% and 10.68% of the phenotypic variation in stem length were determined and two QTLs with minor effect were detected on LG F and N. Identification of QTLs for stem length and mapping individual locus should facilitate to describe genetic mechanisms for stem length in different population. SSR markers tightly linked to QTLs for stem length allow to accelerate the elimination of deleterious genes and selection for desirable recombinants at early stage in crop breeding programs.