• Title/Summary/Keyword: PCR polymorphic

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GENETIC POLYMORPHISMS OF THE GLUTATHIONE S-TRANSFERASE AND CYP1A1 GENES IN KOREAN ORAL SQUAMOUS CELL CARCINOMA (한국인 구강 편평세포암에서 Glutathione S-transferase와 CYP1A1 유전자의 다형성)

  • Cha, In-Ho;Kwon, Jong-Jin;Park, Kwang-Kyun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.28 no.5
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    • pp.364-371
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    • 2002
  • Many chemical compopunds are converted into reactive electrophilic metabolites by the oxidative(Phase I) enzymes, which are mainly cytochrome P-450 enzyme(CYPs). Phase II conjugating enzymes, such as glutathione S-transferase(GST), usually act as inactivation of enzymes. Genetic polymorphisms have been found to be associated with increased susceptibility to cancer of the lung, bladder, breast and colorectal. Many of the polymorphic genes of carcinogen metabolism show considerably different type of cancer among different ethnic groups as well as individuals within the same group. The aim of this study is (1) to establish the frequencies of genetic polymorphisms of GSTM1 and CYP1A1 in Korean oral squamous cell carcinoma(SCC), (2) to associate oral SCC with the risk of these genetic polymorphisms. The genetic polymorphisms of the GSTM1 and the CYP1A1 genes among 50 Korean oral SCC were analyzed using polymerase chain reaction(PCR). The results suggest that the homozygote and the mutant type of CYP1A1 MspI polymorphisms may be associated with genetic susceptibility to oral SCC in Korean. A combination of the GSTM1 null type with the homozygote(m1/m1), and the mutant(m2/m2) type of CYP1A1 MspI polymorphisms showed a relatively high risk of oral SCC in Korean. In the smoking group, the GSTM1 wild genotype may be the high risk factor of oral SCC in Korean. These data coincide with the hypothesis which states that different susceptibility to cancer of genetic polymorphisms exist among different ethnic group and different types of human cancer.

Distribution of the 9-bp Deletion in Coll/$tRNA^{Lys}$ Intergenic Region of Mitochondrial DNA is Relatively Homogeneous in East Asian Populations

  • Hong, Seong-Su;Horai, Satoshi;Lee, Chung-Choo
    • Animal cells and systems
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    • v.2 no.2
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    • pp.259-267
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    • 1998
  • A deletion of one out of the two copies of 9-bp repeat sequence (CCCCCTCTA), between the cytochrome oxidase II and Iysine tranfer RNA (COII/$tRNA^{Lys}$) genes in human mitochondrial DNA (mtDNA) has been used as a polymorphic anthropological marker for people of east Asian origin, and to lesser extent, Pacific and African populations. We searched for the 9-bp deletion of the intergenic COII/$tRNA^{Lys}$ Lys region in two Korean populations (175 from Seoul and 38 from Cheju) and examine the distibution of this deletion in world populations. The 9-bp deletion was detected directly by electrophoresis of the polymerase chain reaction (PCR)-amplified nucleotide(nt) 8211-8310 mtDNA fragment. The frequencies of the 9-bp deletion were significantly different between the Seoul (16%) and Cheju (8%) populations. Examination of data from the world populations suggests a geographic gradient. The frequency reaches its highest values in some Pacific island populations and decreases along the southeast Asia-Siberia transect. In spite of this geographic gradient, Mongoloid populations including Korean, Chinese, Japanese, and Mongolian populations were relatively homo-geneous with regard to the 9-bp deletion type of the intergenic COII/$tRNA^{Lys}$ region. These results indicate Koreans are genetically related to northeast Asian populations, and have a maternal mongoloid ancestry. Therefore, the 9-bp deletion of the intergenic COII/$tRNA^{Lys}$ region will provide significant information to elucidate the historical patterns of migration of the Mongoloids.

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Genetic Diversity and Phylogenetic Relationship of Genus Phyllostachys by ISSR Markers (ISSR 분자 마커를 이용한 왕대속 대나무의 유전적 다양성 및 계통 관계)

  • Lee, Song-Jin;Huh, Man-Kyu;Huh, Hong-Wook
    • Journal of Life Science
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    • v.17 no.11
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    • pp.1482-1487
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    • 2007
  • Four taxa of the genus Phyllostachys were analysed with PCR-based molecular inter-simple sequence repeat (ISSR) determine markers to determine their phylogenetic relationships. Many species of this genus are regarded as economical and ecologically important in the world. With the nine primers screened, ISSR assay generated a total of 64 reproducible bands. Analysis of ISSR from individual plants of genus Phyllostachys resulted in 43 polymorphic bands with 70.49%. When species were grouped by four taxa, within group diversity was 0.092 (Hs), while among group diversify was 0.499 (Gst) on a per locus basis. The estimated gene flow (Nm) between the pairs of species was 0.559. Hence, we can expect weak or low gene flow among species. Total mean genetic diversity ($H_T$) for four taxa was 0.291. The ISSR data allowed us to resolve well-supported clades in the genus Phyllostachys. The four taxa of the genus Phyllostachys analyzed were distinctly related to a monophyletic.

Vegetative Growth and Phylogenetic Relationship of Commercially Cultivated Strains of Pleurotus eryngii based on ITS sequence and RAPD

  • Alam, Nuhu;Shim, Mi-Ja;Lee, Min-Woong;Shin, Pyung-Gyun;Yoo, Young-Bok;Lee, Tae-Soo
    • Mycobiology
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    • v.37 no.4
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    • pp.258-266
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    • 2009
  • Pleurotus eryngii, known as king oyster mushroom has been widely used for nutritional and medicinal purposes. This study was initiated to screen the suitable conditions for mycelial growth and to determine the phylogenetic relationship of the selected strains. Optimal mycelial growth was observed at $30{^{\circ}C}$ and minimum mycelial growth observed at $10{^{\circ}C}$. This mushroom tolerates a broad pH range for mycelial growth, with most favorable growth observed at pH 6. Results also indicated that glucose peptone, yeast malt extract and mushroom complete media were favorable growth media, while Hennerberg and Hoppkins media were unfavorable. Dextrin was the best and xylose the least effective carbon sources. Results revealed that inorganic nitrogen sources were less effective than organic sources for the mycelial growth of P. eryngii. Investigation of genetic diversity is necessary to identify the strains. The ITS region of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 214 to 222 bp and 145 to 236 bp, respectively. The sequence of ITS2 was more variable than that of ITS1, and the 5.8S sequences were identical. A phylogenetic tree based on the ITS region sequences indicated that selected strains could be classified into six clusters. Fourteen IUM and ATCC- 90212 strains were also analyzed by RAPD with 20 arbitrary primers. Fourteen of these primers were efficiently amplified the genomic DNA. The number of amplified bands varied with the primers and strains, with polymorphic fragments in the range from 0.2 to 2.3 kb.

Diversity and Inheritance of AFLP Markers in Wild and Cultivated Soybeans (AFLP marker를 이용한 콩의 유전적 다양성과 유전분리 분석)

  • 김용호;윤홍태
    • Korean Journal of Plant Resources
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    • v.17 no.3
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    • pp.265-271
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    • 2004
  • Genetic variation is the basis of crop improvement. Limited genetic diversity in a crop species may restrict the amount of genetic improvement that can be achieved through plant breeding. Soybean is one of the world's most important crops. A potential source of genetic variability for the cultivated soybean is the wild species G. soja Sieb. & Zucc. Amplified fragment length polymorphism (AFLP) analysis is a PCR-based technique, which can detect a 10-fold greater nubmer of loci than other DNA marker analysis. Twenty cultivated soybeans and two-hundred wild soybeans were used to determine genetic vatiations by AFLPs and evaluate the usefulness of AFLPs as DNA markers. Six-hundred and ten fragments were detected with an average of 56 AFLP fragments produced per primer in a total of 11 AFLP primer pairs. The number of polymorphic loci detected per primer ranged from 7 to 20 and the polymorphism was greater in wild than in cultivated soybean. F$_2$ segregation analysis of four AFLP fragments in combination of Hwaeomputkong ${\times}$ PI 417479 indicated that they segregate as stable Mendelian loci with 3 : 1. This results strongly suggest that the AFLP analysis is a good technique for the detection of genetic polymorphism in a wide plant species.

Genetic diversity and relationship of Korean chicken breeds using 12 microsatellite markers

  • Kim, Yesong;Yun, Ji Hye;Moon, Seon Jeong;Seong, Jiyeon;Kong, Hong Sik
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.3
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    • pp.154-161
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    • 2021
  • A number of Korean Chicken breeds were registered in Domestic Animal Diversity Information System (DAD-IS, http://dad.fao.org/) of the Food and Agriculture Organization (FAO). Evaluation of genetic diversity and relationship of local breeds is an important factor towards the identification of unique and valuable genetic resources. Therefore, this study aimed to analysis the genetic diversity and relationship of 22 Korean Chicken breeds using 12 microsatellite (MS) markers. The mean number of alleles for each variety was 5.52, ranging from a 3.75 (Leghorn F; NF) to a 7.0 (Ross). The most diverse breed was the Hanhyup3 (HCC), which had the highest expected heterozygosity (HExp) (0.754) and polymorphic information content (PIC) (0.711). The NF was the least diverse population, having the lowest HExp (0.467) and PIC (0.413). As a result of the principal coordinates analysis (PCoA) and factorial correspondence analysis (FCA) confirmed that Hy-line Brown (HL) and Lohmann Brown (LO) are very close to each other and that Leghorn and Rhode Island Red (RIR) are clearly distinguished from other groups. Thus, the reliability and power of identification using 12 types of MS markers were improved, and the genetic diversity and probability of individual discrimination were confirmed through statistical analysis. This study is expected to be used as basic data for the identification of Korean chicken breeds, and our results indicated that these multiplex PCR marker sets will have considerable applications in population genetic structure analysis.

Marker Assisted Development and Characterization of Beta-Carotene Rice

  • Yang, Paul;Song, Mi-Hee;Ha, Sun-Hwa;Kim, Jae-Kwang;Park, Jong-Seok;Ahn, Sang-Nag
    • Korean Journal of Breeding Science
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    • v.43 no.5
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    • pp.360-367
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    • 2011
  • Beta-carotene producing transformants were produced in the background of 'Nagdongbyeo', a Japonica rice cultivar. Introgression of the carotenoid locus in the transformant, PAC4-2 into the elite cultivar 'Ilpumbyeo' was started. To initiate a backcrossing program, we surveyed 220 SSR markers and found that 38% of them were polymorphic between 'Ilpumbyeo' as a recurrent parent and the PAC4-2 as a recipient parent. The selection strategy comprising foreground and background selection was employed. First, foreground selection was practiced in $BC_1$, $BC_2$, and $BC_3$ generations using the transgene specific PCR-based marker in addition to visual scoring of the seed color. Marker-based background selection combined with phenotypic selection was employed from $BC_3F_2$ to $BC_3F_4$ generations. Blast search indicated that the transgene PAC4-2 was located between SSR markers, RM6 and RM482. 240 $BC_3F_3$ and 63 $BC_3F_4$ lines were evaluated for four agronomic traits including days to heading. Most of the lines were similar to Ilpumbyeo in agronomic traits evaluated. The percentage of PAC4-2 genome ranged from 4% to 21% with a mean of 12.5%, which was higher than the expected for an unselected $BC_3$ backcross population. This could be explained by the fact that two genes for beta-carotene and the stripe virus resistance were targeted in this study. We selected 10 representative $BC_3F_5$ lines from 63 $BC_3F_4$ lines based on agronomic traits and carotenoids content. The selection strategy would be appropriate for the introgression of beta-carotene gene in a breeding program.

Genetic diversity of grapevine (Vitis vinifera L.) as revealed by ISSR markers

  • Basheer-Salimia, Rezq;Mujahed, Arwa
    • Journal of Plant Biotechnology
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    • v.46 no.1
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    • pp.1-8
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    • 2019
  • The main goal of this study was to determine the genetic diversity among 36 grape cultivars grown in Palestine by using ISSR-polymerase chain reaction (PCR) fingerprints. Among the tested primers, 17 produced reasonable amplification products with high intensity and pattern stability. A total of 57 DNA fragments (loci) separated by electrophoresis on agarose gels were detected and they ranged in size, from 150 to 900 bp. Out of these fragments, 55 (88%) were polymorphic and 2 (3.5%) monomorphic. Our results also revealed an average of 3.1 loci per primer. A minimum of 1 and maximum of 10 DNA fragments were obtained (S-17, #820 and #841) and (S-31) primers, respectively. Therefore, the later primer (S-31) is considered to be the most powerful primer among the tested ones. The genetic distance matrix showed an average distance range of between 0.05 and 0.76. The maximum genetic distance value of 0.76 (24% similarity) was exhibited between the (Shami and Marawi.Hamadani.Adi) as well as (Bairuti and Marawi.Hamadani.Adi) genotypes. On the other hand, the lowest genetic distance of 0.05 (95% similarity) was exhibited between (Jandali.Tawel.Mofarad and Jandali. Kurawi.Mlzlz) along with (Shami.Aswad and Shami.mtartash. mlwn) genotypes. Furthermore, the UPGMA dendrogram generally clusters the grape cultivars into eight major clusters in addition to an isolated genotype. Based on these figures, the cultivars tested in this study could be characterized by large divergence at the DNA level. This is taking the assumption that our region has a very rich and varied clonal grape genetic structure.

Comparative mitogenomics of Pleurotus ostreatus Gonji7ho and its cap color mutant

  • Oh, Min-Ji;Na, Kyeong Sook;Jung, Hwa Jin;Lee, Young Kuk;Ryu, Jae-San
    • Journal of Mushroom
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    • v.20 no.2
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    • pp.43-49
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    • 2022
  • Pleurotus ostreatus is a globally cultivated mushroom crop. Cap color is a quality factor in P. ostreatus. However, cap color can spontaneously mutate, degrading the quality of the mushroom on the market. Early detection and removal of mutant strains is the best way to maintain the commercial value of the crop. To detect the cap color mutant Gonji7ho, molecular markers were developed based on insertion/deletions (InDels) derived from the comparison of mitogenomes of Gonji7ho and Gonji7hoM mushrooms. Sequencing, assembly, and comparative analysis of the two mitogenomes revealed genome sizes of 73,212 bp and 72,576 bp with 61 and 57 genes or open reading frames (ORFs) in P. ostreatus Gonji7ho and Gonji7hoM, respectively. Fourteen core protein-encoding genes, two rRNA, and 24 tRNA with some OFRs were predicted. Of the 61 genes or OFRs in the wild type, dpo, rpo, and two orf139 were missing (or remnant) in the mutant strain. Molecular markers were developed based on the sequence variations (InDels) between the two mitogenomes. Six polymorphic molecular markers could detect the mutated mitochondria by PCR. These results provide basic knowledge of the mitogenomes of wild-type and mutant P. ostreatus, and can be applied to discriminate mutated mitochondria.

Analysis of phylogenetic relationship among Korean Pinellia Tenore (Araceae) using RAPD markers (RAPD markers에 의한 한국산 반하속 식물의 유연관계 분석)

  • Tae, Kyoung-Hwan;Kim, Dong-Kap;Kim, Joo-Hwan
    • Korean Journal of Plant Taxonomy
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    • v.35 no.3
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    • pp.161-174
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    • 2005
  • In order to presume the relationships between two species of P. ternata and P. tripartia, and their populations of the Korean Pinellia, RAPD analysis was performed. The length of the amplified DNA fragments ranged from 300 to 2,500bp. Seventy scorable RAPD makers were found from the PCR reactions with 7 random oligoprimers and were analyzed by Nei-Li's genetic coefficient. Also, some regional groups instead of same taxa were clustered from the phenogram of UPGMA analysis and NJ tree. Populations within each species were clustered at low genetic distance, there had the closed relationship. According to the regional individuals, Pinellia ternata was showed the variation pattern of morphological (leaf shape and flower color) and cytological characters(somatic chromosome numbers). So we suggested to difference of characteristic variety based on variety of habitat. According to this study, new species (Pinellia sp.) was affiliated with Pinellia and had the closest relationship with Hallasan and Japan population. The RAPD data was very useful to define the genetic variation and to discuss the relationships among the intraspecific taxa and their populations of the Korean Pinellia.