• Title/Summary/Keyword: marker selection

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Optimized Protocols for Efficient Plant Regeneration and Gene Transfer in Pepper (Capsicum annuum L.)

  • Mihalka, Virag;Fari, Miklos;Szasz, Attila;Balazs, Ervin;Nagy, Istvan
    • Journal of Plant Biotechnology
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    • v.2 no.3
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    • pp.143-149
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    • 2000
  • An Efficient in vitro regeneration system and an optimized Agrobacterium mediated transformation protocol are described, based on the use of young seedling cotyledons of Capsicum annuum L. Optimal regeneration efficiency can be obtained by cultivating cotyledon explants on media containing 4 mg/L benzyladenine and 0.1 mg/L indolacetic acid. The effect of antibiotics used to eliminate Agrobacteria, as well as the toxic level of some generally used selection agents (kanamycin, geneticin, hygromycin, phosphinotricin and methotrexate) in regenerating pepper tissues were determined. To enable the comparison of different selection markers in identical vector background, a set of binary vectors containing the marker genes for NPTII, HPT, DHFR and BAR respectively, as well as the CaMV 35S promoter/enhancer-GUS chimaeric gene was constructed and introduced into four different Agrobacterium host strains.

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Characteristics of the Plasmid pCS100 Containing Nisin Resistant Gene from Lactococcus lactis subsp. lactis ATCC7962. (Lactococcus lactis subsp. lactis ATCC 7962의 nisin 저항성 유전자를 포함하는 plasmid pCS100의 특성규명)

  • 송종효;이형주;김정환;정대균
    • Microbiology and Biotechnology Letters
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    • v.26 no.6
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    • pp.562-565
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    • 1998
  • Nisin-producing and nisin resistant L. lactis subsp. lactis ATCC7962 harbored six plasmids. To find a plasmid containing a nisin resistant gene, these plasmids were transformed into L lactis LM0230 of plasmid-free and nisin sensitive strain. After screening on nisin selection media containing nisin (150 $\mu\textrm{g}$/$m\ell$), several nisin resistant transformants were obtained and the level of nisin resistance was very similar to that of wild type L lactis subsp. lactis ATCC7962. A 26.5 kb plasmid, named as pCS100, which confers resistance to nisin, was identified in transformants. The pCS100 was digested with EcoRI and Southern blot hybridization was done with nisI probe to localize the nisin resistant gene. A 4 kb EcoRI fragment showed a strong positive signal, and it was cloned into pBluescript for the potential selection marker.

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Improved Transformation of the Filamentous Fungus Aspergillus niger Using Agrobacterium tumefaciens

  • Park, Seung-Moon
    • Mycobiology
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    • v.29 no.3
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    • pp.132-134
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    • 2001
  • Since it is known that Agrobacterium tumefaciens, which has long been used to transform plants, can transfer the T-DNA to yeast Saccharomyces cerevisiae during tumourigenesis, a variety of fungi were subjected to transformation to improve their transformation frequency. In this study, I report the A. tumefaciens-mediated transformation of filamentous fungus Aspergillus niger. Transfer of the binary vector pBIN9-Hg, containing the bacterial hygromycin B phosphotransferase gene under the control of the Aspergillus nidulans trpC promoter and terminator as a selectable marker, led to the selection of $50{\sim}100$ hygromycin B-resistant transformants per $1{\times}10^7$ conidia of A. niger. This efficiency is improved $10{\sim}20$ fold more than reported elsewhere. In order to avoid the difficulties in selection transformant from the over-growing non-transformant, I used top agar containing 900 ${\mu}g/ml$ of hygromycin. Genomic PCR and Southern analysis showed that all transformants contained single T-DNA insert per fungal genome. This technique offers an easier and more efficient method than that of using protoplast.

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Implementation of genomic selection in Hanwoo breeding program (유전체정보활용 한우개량효율 증진)

  • Lee, Seung Hwan;Cho, Yong Min;Lee, Jun Heon;Oh, Seong Jong
    • Korean Journal of Agricultural Science
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    • v.42 no.4
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    • pp.397-406
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    • 2015
  • Quantitative traits are mostly controlled by a large number of genes. Some of these genes tend to have a large effect on quantitative traits in cattle and are known as major genes primarily located at quantitative trait loci (QTL). The genetic merit of animals can be estimated by genomic selection, which uses genome-wide SNP panels and statistical methods that capture the effects of large numbers of SNPs simultaneously. In practice, the accuracy of genomic predictions will depend on the size and structure of reference and training population, the effective population size, the density of marker and the genetic architecture of the traits such as number of loci affecting the traits and distribution of their effects. In this review, we focus on the structure of Hanwoo reference and training population in terms of accuracy of genomic prediction and we then discuss of genetic architecture of intramuscular fat(IMF) and marbling score(MS) to estimate genomic breeding value in real small size of reference population.

RAPD Pattern of Ginseng(Panax ginseng C.A. Meyer) Lines Containing High Level of Ginsenoside

  • Kang, Tae-Jin;Kim, Se-Young;Rho, Yeong-Deok;Deok-Chun
    • Plant Resources
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    • v.6 no.3
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    • pp.170-174
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    • 2003
  • The important component for medical effect in ginseng is ginsenoside. Korea Ginseng & Tobacco Research Institute contains approximately 200 lines produced by inbred selection. It is assumed that ginseng lines containing high level of ginsenoside should be included in those lines. Besides, new breeding methods such as cell line selection in vitro and hairy root were recently developed. Therefore, this study was carried out to detect genes related to ginsenoside, and to use it for selection marker to select and distribute lines containing high level of ginsenoside. DNA was extracted from both ginseng roots and hairy roots, and the difference between the line containing high ginsenoside(KG101) and normal ginsenoside(KG103) were analysed. As a result, 28 out of 36 primers showed bands, and many primers showed band difference between ginseng lines. It is considered that the bands should be analysed using DNA sequence comparison to check if those are related to ginsenoside. In case of hairy roots of ginseng, almost no differences were found between two lines.

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Food-Grade Expression and Secretion Systems in Lactococcus

  • Jeong, Do-Won;Hwang, Eun-Sun;Lee, Hyong-Joo
    • Food Science and Biotechnology
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    • v.15 no.4
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    • pp.485-493
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    • 2006
  • Lactococcus species are noninvasive and nonpathogenic microorganisms that are widely used in industrial food fermentation and as well-known probiotics. They have been modified by traditional methods and genetic engineering to produce useful food-grade materials. The application of genetically modified lactococci in the food industry requires their genetic elements to be safe and stable from integration with endogenous food microorganisms. In addition, selection for antibiotic-resistance genes should be avoided. Several expression and secretion signals have been developed for the production and secretion of useful proteins in lactococci. Food-grade systems composed of genetic elements from lactic acid bacteria have been developed. Recent developments in this area have focused on food-grade selection markers, stabilization, and integration strategies, as well as approaches for controlled gene expression and secretion of foreign proteins. This paper reviews the expression and secretion signals available in lactococci and the development of food-grade markers, food-grade cloning vectors, and integrative food-grade systems.

Effects of Selection by Serum IGF-I Concentration in Korean Native Ogol Chicken

  • Kim, D. H.;Kim, M. H.;W. J. Kang;D. S. Seo;Y. Ko
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.20-20
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    • 2003
  • Phenotypic characteristics and genetic markers in livestock have been utilized for improvement of the economic traits including egg productivity. Korean Native Ogol Chicken (KNOC) has low egg productivity compared to White Leghorn. Therefore, in this study, serum IGF-I concentration and number of egg production were used as selection markers to improve egg productivity. KNOCs were divided into three groups showing high IGF-I concentration (IGF-I high), high egg production (EP high), and IGF-I/EP high groups. Blood was collected every 10 weeks, and serum concentrations of IGF-I, estradiol (E2), and progesterone (P4) were measured by radioimmunoassay. In comparison of three groups in each generation, the highest increment of egg production was detected in the IGF-I/EP high group from 20 weeks till 40 weeks, and the IGF-I high group also showed the significant increment of egg production after 50 weeks. Interestingly, there were the increase of egg production and decrease of periods in sexual maturity in the second and third generation selected by serum IGF-I concentration, while egg weight and body weight decreased during experimental period. In conclusion, the present study suggest the possibility of IGF-I as a selection marker to improve the egg productivity of KNOC.

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Effects of Single Nucleotide Polymorphism Marker Density on Haplotype Block Partition

  • Kim, Sun Ah;Yoo, Yun Joo
    • Genomics & Informatics
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    • v.14 no.4
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    • pp.196-204
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    • 2016
  • Many researchers have found that one of the most important characteristics of the structure of linkage disequilibrium is that the human genome can be divided into non-overlapping block partitions in which only a small number of haplotypes are observed. The location and distribution of haplotype blocks can be seen as a population property influenced by population genetic events such as selection, mutation, recombination and population structure. In this study, we investigate the effects of the density of markers relative to the full set of all polymorphisms in the region on the results of haplotype partitioning for five popular haplotype block partition methods: three methods in Haploview (confidence interval, four gamete test, and solid spine), MIG++ implemented in PLINK 1.9 and S-MIG++. We used several experimental datasets obtained by sampling subsets of single nucleotide polymorphism (SNP) markers of chromosome 22 region in the 1000 Genomes Project data and also the HapMap phase 3 data to compare the results of haplotype block partitions by five methods. With decreasing sampling ratio down to 20% of the original SNP markers, the total number of haplotype blocks decreases and the length of haplotype blocks increases for all algorithms. When we examined the marker-independence of the haplotype block locations constructed from the datasets of different density, the results using below 50% of the entire SNP markers were very different from the results using the entire SNP markers. We conclude that the haplotype block construction results should be used and interpreted carefully depending on the selection of markers and the purpose of the study.