• Title/Summary/Keyword: ITS-RFLPs

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Varietal Classification of Barley by Isozymes and Restriction Fragment Length Polymorphisms (RFLPs) (동위효소 분석과 제한효소 단편 다형화현상을 이용한 보리 품종의 분류)

  • Jin, Byung-Soon;Park, Ro-Dong;Eun, Moo-Young;Lee, Eun-Seop
    • Applied Biological Chemistry
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    • v.36 no.3
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    • pp.139-145
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    • 1993
  • The methods of isoelectric focusing of 4 isozymes in polyacrylamide horizontal slab gels and restriction fragment length polymorphisms (RFLPs) were applied to characterize the biochemical phenotypes of 19 cultivars of barley. Among 19 barley cultivars screened, 7 esterase, 3 phosphoglucose isomerase, 4 peroxidase and 2 alcohol dehydrogenase isozyme phenotypes were distinguished by isoelectric focusing. When purified DNA of each cultivar was digested with restriction enzyme EcoRV and analyzed its RFLPs with barley DNA markers pMSU 51 or pMSU 71, two distinct RFLP patterns were shown. Based on the four isozymes and two RELP polymorphisms, 19 cultivars of barley were classified into 13 biochemical phenotypes. Phylogenetical relationships among 13 biochemical phenotypes classified were determined using Nei's F-statistics and the phylogenetic dendrogram was constructed.

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PCR-DGGE and PCR-RFLP Analyses of the Internal Trascribed Spacer(ITS) of Ribosomal DNA in the Genus Rhizopus

  • Park, You-jung;Park, Young-Keel;Min, Byung-Re
    • Journal of Microbiology
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    • v.41 no.2
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    • pp.157-160
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    • 2003
  • To estimate genetic relationships within the genus Rhizopus, genetic variations in 20 strains were investigated by DGGE and PCR-RFLP of rDNA ITS region (ITSI, ITS2,5.8S). The size of the amplified products showed the interspecific polymorphisms, 650 bp,700 bp, and 900 bp. The DGGE approach allowed the separation of PCR amplicons of the same length according to their sequence variations. When the rDNA ITS region was digested with six restriction enzymes, 20 strains were classified into five RFLP haplotypes. The range of similarity between the 20 strains by PCR-RFLP was 42.3-100%. Based on the results of DGGE aud PCR-RFLP, the 20 strains were divided into four groups, R. oryzae, R. stolonifer, R. microsporus and R. homothallicus. Further study of R. homothallicus is required.

Molecular Markers and Their Application in Mulberry Breeding

  • Vijayan, Kunjupillai
    • International Journal of Industrial Entomology and Biomaterials
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    • v.15 no.2
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    • pp.145-155
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    • 2007
  • Mulberry (Morus spp.) is an economically important tree crop being cultivated in India, China and other sericulturally important countries for its foliage to feed the silk producing insect Bombyx mori L. Genetic improvements of mulberry lag behind to the same in many other economically less important crops due to the complexity of its genetics, the breeding behavior, and the lack of basic information on factors governing important agronomic traits. In this review, the general usage and advantages of different molecular markers including isoenzymes, RFLPs, RAPDs, ISSRs, SSRs, AFLPs and SNPs are described to enlighten their applicability in mulberry genetic improvement programs. Application of DNA markers in germplasm characterization, construction of genetic linkage maps, QTL identification and in marker-assisted selection was also described along with its present status and future prospects.

Identification and Phylogenetic Relationship of Dermatophytes Based on RFLP Analysis and Nucleotide Sequence of Internal Transcribed Spacer (ITS)1 in Nuclear Ribosome DNA (ITS-RFLP와 ITS1 염기서열 분석에 의한 피부사상균의 동정과 계통적 유연관계)

  • Choi, Yeon-Hwa;Lee, Yeong-Seon;Yoo, Jae-Il;Kim, Bong-Su
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.49-60
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    • 2000
  • ITSI-5.8S-ITSII rDNA region was amplified from the reference strains and clinical isolates with ITS1 and ITS4 primers. These primers amplified DNA fragments of 550 bp in Microsporum audouinii and Trichophyton violaceum, 700 bp in Microsporum gypseum, Trichophyton mentagrophytes, Trichophyton rubrum, and Trichophyton tonsurans, and 750 bp in Microsporum ferreugineum and Microsporum canis. The restriction enzyme patterns of PCR products digested with 13 restriction enzyme including PstI were distint among the genera, whereas identical in the same species. Examination of the ITS (Internal Transcribed Spacers)1 nucleotide sequence revealed that there was the genetic difference in each genera and species. Phylogenetic relationship among each species showed that the Trichophyton mentagrophytes was more closely related Trichophyton tonsurans than Trichophyton rubrum, and Microsporum gypseum was less related than Microsporum spp..

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Genetic Variation of H-FABP Gene and Association with Intramuscular Fat Content in Laiwu Black and Four Western Pig Breeds

  • Zeng, Y.Q.;Wang, G.L.;Wang, C.F.;Wei, S.D.;Wu, Y.;Wang, L.Y.;Wang, H.;Yang, H.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.1
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    • pp.13-16
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    • 2005
  • This study was performed to detect genetic variation of the heart fatty acid-binding protein (H-FABP) gene by PCRRFLPs approach and its association with intramuscular fat (IMF) content. Data from 223 individuals, including one Chinese native pig breed and four western pig breeds, were analyzed. The results showed that for the H-FABP gene, there was one polymorphic HinfI site in the 5'-upstream region, whereas there were one HaeIII and one HinfI (marked as $HinfI^*$) polymorphic site in the second intron, respectively. The three PCR-RFLPs were present in all breeds tested. The allele frequencies, however, revealed significant differences between them (p<0.05). Furthermore, the allele frequency distribution of HinfI in the Laiwu Black and that of $HinfI^*$ in the Hampshire breed were at disequilibrium, which might be the result of selective breeding. Results also indicated that for HinfI, HaeIII and $HinfI^*$ HFABP RFLP, significant (p<0.05) contrasts of 0.78%, -0.69% and 0.72% were detected in the least square means of IMF content between the homozygous genotype HH and hh, DD and dd, BB and bb classes, respectively. It implied that the HHddBB genotype had the highest IMF content in this experimental population and these H-FABP RFLPs could serve, to some extent, as genetic markers for use in improvement of IMF content.

FSHR Gene Mutation and Its Effect on Litter Size in Pigs

  • Zhang, Shujun;Xiong, Yuanzhu;Den, Changyan;Xiao, Senmu;Xu, Jianxiang;Xia, Yu;Liu, Xiaohua;Wang, Chunfang;Sun, Shulin
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.2
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    • pp.179-182
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    • 2004
  • The polymorphism of the locus FSHRB in intron6-Exon7 of FSHR gene was investigated by PCR-RFLPs in Erhualian, Large White and Landrace${\times}$Large White; The association of polymorphism and litter size was analyzed by using SAS. The results showed that 1) the polymorphism of the locus FSHRB was significantly associated with litter size; 2) the total born number (TBN) and number born alive (NBA) of the sows with BB genotype were increased (p<0.05) with additive effects of 1.02-1.42 and 1.04-1.27 pigs per litter, respectively; 3) among the sows with genotype AA, AB or BB, there was an insignificant difference in born weight and weaning weight. This gene may be an effective potential tool used in conjunction with traditional selection methods.

A Molecular Marker Specific to Metarhizium anisopliae var. majus

  • YOON, CHEOL-SIK;GI HO SUNG;JAE MO SUNG;JAEANG OON LEE
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.334-339
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    • 1999
  • More innovative molecular markers were investigated for rapid and consistent differentiation of Metarhizium anisopliae var. majus from M. anisopliae var. anisopliae. A total of 28 isolates were obtained from various countries and hosts: 13 isolates of M. anisopliae var. anisopliae, 12 isolates of M. anisopliae var. majus, and 3 isolates of M. anisopliae collected in Korea. This study involved restriction enzyme digestions of a PCR product amplified from nuclear internally transcribed spacer (ITS) and a portion of the 28S rDNA regions. Among 11 different restriction enzymes used in this study, MboⅠ digestion particularly produced a restriction pattern that had characteristics of M. anisopliae var. majus. This restriction pattern was consistent in all isolates of M. anisopliae var. majus regardless of their geographic origins and insect hosts. Mapping experiments revealed that MboⅠ sites of M. anisopliae var. majus are identical to those of M. anisopliae var. anisopliae with an exception for the presence of an additional site in the PCR product. Results from this study provide an additional method for identification and differentiation of isolates of these two varieties of M. anisopliae for use in the field and laboratory experiments.

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DNA Polymorphism in SLC11A1 Gene and its Association with Brucellosis Resistance in Indian Zebu (Bos indicus) and Crossbred (Bos indicus×Bos taurus) Cattle

  • Kumar, Nishant;Ganguly, Indrajit;Singh, Rajendra;Deb, Sitangsu M.;Kumar, Subodh;Sharma, Arjava;Mitra, Abhijit
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.7
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    • pp.898-904
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    • 2011
  • The PCR- restriction fragment length polymorphism (RFLP) in and around TM4 of SLC11A1 gene and its association with the incidences of brucellosis in Hariana breed (Bos indicus) and Holstein Friesian crossbred (Bos indicus${\times}$Bos taurus) cattle was examined. A fragment of 954 bp encoding the TM4 was amplified, and RFLP was identified by digestion of the amplicon independently with AluI and TaqI. The amplicon (GenBank Acc. No. AY338470 and AY338471) comprised of a part of exon V (<59 bp) and VII (62>), and entire intron 5 (423 bp), exon VI (71 bp) and intron 6 (339 bp). Digestion with AluI revealed the presence of two alleles viz, A (281, 255, 79 and 51 bp) and B (541, 255, 79 and 51 bp). The frequency of A allele was estimated as 0.80 and 0.73 in Hariana and crossbred cattle, respectively. Due to presence of a polymorphic TaqI site at intron 5, two alleles: T (552 and 402 bp) and Q (231, 321 and 402 bp) were identified. The frequency of T allele was estimated as 0.96 and 0.97, respectively. For association study, on the basis of serological tests and history of abortion, the animals were grouped into "affected" and "non-affected". However, no association could be established with the observed RFLPs.

First Note of Hypoxylon truncatum sensu Miller in Korea (국내에서 발견된 Hypoxylon truncatum sensu Miller의 형태 및 유전적 특징)

  • Yoon, Cheol-Sik;Chang, Hyun-You;Sung, Gi-Ho
    • The Korean Journal of Mycology
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    • v.25 no.3 s.82
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    • pp.161-166
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    • 1997
  • Occurrence of Hypoxylon truncatum, known as a symbiont of white jelly mushroom, was first noted and described in Korea. Two stromatal forms reported in North American collections of H. truncatum sensu Miller were also observed in Korean collections. Based on evidences from other studies, polymorphic patterns seen in six enzyme digestions of nuclear Internal Transcribed Spacer (ITS) regions of Korean isolates indicated that the two stromatal forms were genetically distinct. Because there was a clear association of stromatal morphology with genetic differences, the different stromatal forms might be different species. In addition, clear species concept on the species H. truncatum would provide aids in selecting proper strain for cultivation of white jelly mushrooms.

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A study of Association of the H-FABP RFLP with Economic Traits of Pigs (돼지 H-FABP 유전자의 다형성 및 경제 형질과의 연관성 구명)

  • Choi, B.H.;Kim, T.H.;Lee, J.W.;Cho, Y.M.;Lee, H.Y.;Cho, B.W.;Cheong, I.C.
    • Journal of Animal Science and Technology
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    • v.45 no.5
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    • pp.703-710
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    • 2003
  • The purpose of this study was to detect association between genetic variation and economic trait in the porcine heart type fatty acid-binding protein gene as a candidate gene for the traits related with growth and meat quality in pigs. The H-FABP is a 15-kDa protein expressed in several tissues with high demand for fat metabolism such as cardiac and skeletal muscle and lactating mammary gland. H-FABP is small intracellular protein involved in fatty acid transport from the plasma membrane to the site of $\beta$-oxidation and/or triacylglycerol or phospholipid synthesis. In this study, H-FABP PCR-RFLP was performed in F$_2$ population composed of 214 individuals from an intercross between Korean Native Boars and Landrace sows. PCR products from two primer sets within H-FABP gene were amplified in 850bp and 700bp. Digestion of PCR products with the restriction digestion enzymes HaeⅢ and HinfⅠ, revealed fragment length polymorphisms(RFLPs). The genotype frequencies from H-FABP/HaeⅢ was .29 for genotype DD, .53 for genotype Dd, and .15 for genotype dd, respectively. The genotype frequencies of HH, Hh, and hh from H-FABP/HinfⅠ was .38, .41 and .20, respectively, in the population. Relationships between their genotypes and economic traits were estimated. In H-FABP/HaeⅢ locus, there were specific genotypes(Dd and dd) associated with economic traits such as body weights at 3, 5, 12, and 30 week of age (p〈.05 to .001). The ‘d’ allele was associated with gaining of body weight. In H-FABP/HinfⅠ locus, Genotypes of HH and Hh associated with growth traits such as body weights at 5, 12, and 30 week of age (p〈.05 or p〈.001) and back fat thickness, body fat including abdominal and trimmed fat (p〈.001) and intramuscular fat(p〈.05) The ‘H’ allele was positively associated with gaining of body weight and fatness deposition. In conclusion, a significant association of the H-FABP gene from its genetic variation was found on body weight, intramuscular fat and backfat thickness.