Purpose : Glycogen storage disease type III (GSD-III) is a rare autosomal recessive disorder of glycogen metabolism. The affected enzyme, amylo-1,6-glucosidase, 4-alpha-glucanotransferase (AGL, glycogen debranching enzyme), is responsible for the debranching of the glycogen molecule during catabolism. The disease shows clinical and biochemical heterogeneity, reflecting genotype-phenotype heterogeneity among different patients. In this study, we aim at analyzing mutations of the AGL gene in three unrelated Korean GSD-III patients, and characterizing their clinical and laboratory findings. Methods : We characterized the clinical features of three unrelated Korean GSD-III patients by biochemical, histological and imaging studies. The 35 exons and part of exon-intron boundaries of AGL were analyzed by direct sequencing using genomic DNA extracted from the peripheral leukocytes of patients. Results : Diverse clinical features were observed in these patients including hepatomegaly (all patients), seizures (patient 2), grow th failure (patients 1 and 2), hyperlipidemia (patients 1 and 3), raised transaminase and creatine kinase concentrations (all patients), and mild cardiomyopathy (patient 2). Liver transplantation w as performed in patient 2 due to progressive hepatic fibrosis. A dministration of uncooked corn starch maintained normoglycemia and improved biochemical and growth profiles. DNA sequence analysis revealed mutations in 5 out of 6 alleles. Patient 1 was a compound heterozygote of c.1282 G>A (p.R428K) and c.1306delA (p.S603PfsX6), patient 2 had c.1510_1511insT (p.Y 504L fsX 10), and patient 3 had c.3416 T >C (p.L 1139P) and c.1735+1 G>T (p.Y 538_R578delfsX 4) mutations. A part from the p.R428K mutation, the 4 other substitutions identified w ere nov el. Conclusion : GSD-III patients display variable phenotypic characteristics resembling those of GSD-Ia. Molecular defects in the AGL gene of Korean GSD-III patients are genetically heterogeneous.
Response Surface Methodology (RSM), which is combining with Plackett-Burman design and Box-Behnken experimental design, was applied to optimize the ratios of the nutrient components for carotenoid production by Rhodobacter sphaeroides PS-24 in liquid state fermentation. Nine nutrient ingredients containing yeast extract, sodium acetate, NaCl, $K_2HPO_4$, $MgSO_4$, mono-sodium glutamate, $Na_2CO_3$, $NH_4Cl$ and $CaCl_2$ were finally selected for optimizing the medium composition based on their statistical significance and positive effects on carotenoid yield. Box-Behnken design was employed for further optimization of the selected nutrient components in order to increase carotenoid production. Based on the Box-Behnken assay data, the secondary order coefficient model was set up to investigate the relationship between the carotenoid productivity and nutrient ingredients. The important factors having influence on optimal medium constituents for carotenoid production by Rhodobacter sphaeroides PS-24 were determined as follows: yeast extract 1.23 g, sodium acetate 1 g, $NH_4Cl$ 1.75 g, NaCl 2.5 g, $K_2HPO_4$ 2 g, $MgSO_4$ 1.0 g, mono-sodium glutamate 7.5 g, $Na_2CO_3$ 3.71 g, $NH_4Cl$ 3.5g, $CaCl_2$ 0.01 g, per liter. Maximum carotenoid yield of 18.11 mg/L was measured by confirmatory experiment in liquid culture using 500 L fermenter.
The objectives of this study were to analyze mutant lines of Chinese cabbage ($Brassica$$rapa$ ssp. $pekinensis$) using gene tagging system (plasmid rescue and inverse polymerase chain reaction) and to observe the phenotypic characteristics. Insertional mutants were derived by transferring DNA (T-DNA) of $Agrobacterium$ for functional genomics study in Chinese cabbage. The hypocotyls of Chinese cabbage 'Seoul' were used to obtain transgenic plants with $Agrobacterium$$tumefaciens$ harboring pRCV2 vector. To tag T-DNA from the Chinese cabbage genomic DNA, plasmid rescue and inverse PCR were applied for multiple copies and single copy insertional mutants. These techniques were successfully conducted to Chinese cabbage plant with high efficiency, and as a result, T-DNA of pRCV2 vector showed distinct various integration patterns in the transgenic plant genome. The polyploidy level analysis showed the change in phenotypic characteristics of 13 mutant lines was not due to variation in somatic chromosome number. Compared with wild type, the $T_1$ progenies showed varied phenotypes, such as decreased stamen numbers, larger or smaller flowers, upright growth habit, hairless leaves, chlorosis symptoms, narrow leaves, and deeply serrated leaves. The polyploidy level analysis showed the change in phenotypic characteristics of 13 mutant lines was not due to variation in somatic chromosome number. To tag T-DNA from the Chinese cabbage genomic DNA, plasmid rescue and inverse PCR were applied for multiple copies and single copy insertional mutants. Mutants that showed distinct phenotypic difference compared to wild type with 1 copy of T-DNA by Southern blot analysis, and with 2n = 20 of chromosome number were selected. These selected mutant lines were sequenced flanking DNA, mapped genomic loci, and the genome information of the lines is being recorded in specially developed database.
Carrot (Daucus carota L. var. sativa) is one of the most extensively used vegetable crops in the world and a significant source of nutrient because of its high content of ${\beta}$-carotene, well known as the precursor of vitamin A carotenoid. However, seed-hairs generated and elongated from the epidermal cell of seeds inhibit absorption and germination by various factors such as carotol and so on. Accordingly, mechanical hair removal process is essential before commercialization of carrot seeds. Because of this process, producers will have additional losses such as time consuming, manpower, capital and so on. Furthermore, physical damage of seeds causes irregular germination rate. To overcome such cumbersome weaknesses, new breeding program for developing hairless-seed carrot cultivar has been needed and studies for molecular markers related to seed-hair characteristic is needed for a new breeding program. Therefore, in this study, cDNA libraries from seeds of short-hair seed phenotype CT-SMR 616 OP 659-1 line, hairy-seed phenotype CT-SMR 616 OP 677-14 line and short-hair seed phenotype CT-ATR 615 OP 666-13 line, hairy-seed phenotype CT-ATR 615 OP 671-9 were constructed, respectively. Furthermore, 1,248 ESTs in each line, total 4,992 ESTs were sequenced. As a result, 19 SNP sites and 14 SNP sites in each of 2 combinations were confirmed by analyzing these EST sequences from short-hair and hairy-seed lines. Then we designed SNP primer sets from EST sequences of SNP sites for high resolution melting (HRM) analysis. Designed HRM primers were analyzed using hairy seed phenotype CT-SMR 616 OP 1040 line and short-hair seed phenotype CT-SMR 616 OP 1024, 1025, 1026 lines. One set of HRM primers showed specific difference between the melting curves of hairy and short-hair seed phenotype lines. Based on this result, allele-specific (AS) PCR primers were designed for easier selection between hairy-seed carrot and hairless seed carrot. These results of HRM and AS-PCR are expected to be useful in breeding of hairless seed carrot cultivar as a molecular marker.
Niphon spinosus, Epinephelus bruneus, and Epinephelus septemfasciatus are involved in the Perciformes Order and Serranidae Family. When E. bruneus and E. septemfasciatus are fully grown, the striped pattern on the body gradually disappears. Therefore, morphological classification of adult fishes is quite difficult to identify the differences to N. spinosus. In this study, we investigate the method to differentiate those using PCR. To design the primers, 16S rRNA region of N. spinosus, E. bruneus, and E. septemfasciatus registered in the GeneBank (www.ncbi.nlm.nih.gov) have been used and for the analysis, Bio Edit ver. 7.0.9.0 was used. As a result, it was design NS-003-F/NS-005-R (136 bp), EB-001-F/EB-002-R (181 bp), and ES-001-F/ES-001-R (123 bp) primers for the differentiation of each 3 different fishes. Therefore, the species-specific primer sets would be a useful tool for scientific and speedy differentiation against the illegal distribution for consumer protection.
Park, Ji-Hyun;Kim, Hyoung-Seok;Lee, Gi-Ho;Yu, Jae-Gyung;Park, Young-Doo
Horticultural Science & Technology
/
v.34
no.1
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pp.112-121
/
2016
Development of genetically-modified (GM) crops enables the introduction of new traits to the plant to confer characteristics such as disease resistance, herbicide resistance and human health-promoting bioactivity. Successful commercialization of newly developed GM crops requires stable inheritance of integrated T-DNA and newly introduced traits through the multiple generations. This study was carried out to confirm the stable inheritance of the integrated T-DNA in $T_1$ and $T_2$ transgenic Chinese cabbage (Brassica rapa ssp. pekinensis) that was genetically modified to increase concentrations of phenylethylisothiocyanate (PEITC), which is a potential anti-carcinogenic phytochemical. For this purpose, the IGA 1-3 ($T_1$ generation) and IGA 1-3-5 ($T_2$ generation) lines were selected by PCR and a IGA 1-3 transgenic plant ($T_1$ generation) was analyzed to confirm the T-DNA insertion site in the Chinese cabbage genome by VA-TAIL PCR. The results of this study showed that the introduced T-DNA in IGA 1 line was stably inherited to the next generations without any variations in terms of the structure of the transgenes, and this line also showed the expected transgene function that resulted in increased concentration of PEITC through the multiple generations. Finally, we confirmed the increased QR activity in IGA 1 $T_1$ and $T_2$ transgenic lines, which indicates an enhanced potential anti-carcinogenic bioactivity and its stable inheritance in IGA1 $T_1$ and $T_2$ transgenic lines.
With a broad objective for the development of microbial based fertilizers, a total of 373 strains were isolated from rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass. The efficacy of the isolates to augument overall plant growth was evaluated. After screening for their plant growth promotion and antagonistic properties in vitro efficient strains were further selected. The most efficient strains was characterized by 16S rRNA gene sequences and biochemical techniques and was designated as Bacillus subtilis S37-2. The strains facilitated plant growth and inhibited the plant phathogenic fungi such as Fusarium oxysporum (KACC 40037, Rhizoctonia solani (KACC 40140), and Sclerotinia sclerotiorum (KACC 40457). Pot based bioassay using lettuce as test plant was conducted by inoculating suspension ($10^5$ to $10^8cells\;mL^{-1}$) of B. subtilis S37-2 to the rhizosphere of lettuce cultivated in soil pots. Compared with non-inoculated pots, marked increase in leaf (42.3%) and root mass (48.7%) was observed in the inoculation group where the 50ml of cell mixture ($8.7{\times}10^8cells\;ml^{-1}$) was applied to the rhizosphere of letuce either once or twice. Antagonistic effects of B. subtilis S37-2 strain on S. sclerotiorum (KACC 40457) were tested. All the tested lettuce plants perished after 9 days in treatment containing only S. sclerotiorum, but only 17% of lettuce was perished in the inoculation plot. B. subtilis grew well in the TSB culture medium. The isolates grew better in yeast extracts than peptone and tryptone as nitrogen source. The growth rate was 2~4 times greater at $37^{\circ}C$ as compared with $30^{\circ}C$ incubation temperature. B. subitlis S37-2 produced $0.1{\mu}g\;ml^{-1}$ of IAA (indole 3-acetic acid) in the TSB medium containing L-tryptophan($20mg\;L^{-1}$) in 24 hours.
Choi, Hee Ji;Lee, Soo Bin;Kwon, Hye Mi;Choi, Byung-Ok;Chung, Ki Wha
Journal of Life Science
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v.31
no.10
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pp.913-921
/
2021
Limb-girdle muscular dystrophy (LGMD) which is characterized by progressive muscle weakening of the hip and shoulder shows both dominant and recessive inheritances with many pathogenic genes including TTN. This study performed to identify genetic causes of a male patient with late onset (45 years old) autosomal recessive LGMD and atrial flutter. By application of the whole exome sequencing, we identified bi-allelic variants of TTN gene in the patient. One allele had a single missense variant of [c.24124G>T (p.V8042F)], while the other allele consisted of three missense variants of [c.29222G>C (p.R9741P) + c.67490A>G (p.H22497R) + c.75376C>T (p.R25126C)]. The p.V8042F allele was transmitted from his mother, while the other haplotype allele was putatively transmitted from his father. His two unaffected sons had only the p.R9741P. These variants have been not reported or rarely reported in the public human genome databases (1,000 Genome, gnomAD, and KRGDB). Most variants were located in the highly conserved immunoglobulin or fibronectin domains and were predicted to be pathogenic by the in silico analyses. The TTN giant protein plays a key role in muscle assembly, force transmission at the Z-line, and maintenance of resting tension in the I-band. In conclusion, we think that these bi-allelic compound heterozygous mutations may play a role as the genetic causes of the LGMD phenotype.
Korean 'Sin-Pa' play is a way to examine self-reliance of Korean culture and influence of Japanese culture on the Korean one in the early modern times. Although the 'Sin-Pa' play has been estimated negatively in many aspects, such an estimation can be different depending on the methodology. Therefore, I tried to explain a characteristics of the rise of 'Sin-Pa' play. While making these trials, I made efforts to reappreciate the developing process of Korean 'Sin-Pa' play and its theatrical structure and value. Particularly, I focused on structure of 'Sin-Pa' play in the context of an actual cognition of colonized Korea. Based on the 'Sin-Pa' play's repertoires I found out that one of the representative characteristics of Korean 'Sin-Pa' play is a changing process from orality to literacy. And I made attempts to uncover some ideological functions and their effects of 'Sin-Pa' play, focusing on and whose story line is usually consisted of 'provocation-pangs-defeat' while it interacted with 'provocation-pangs-penalty' of the structure of melodrama under the contemporary cultural conditions. 'Sin-Pa'' play can be considered as a performance mode to accept the Japanese value embedded in the colonized Korea since the 1910s on the one hand and to resist the overwhelming power of western culture imported through Japan on the other hand. In other words, it was closely related with the cultural-field of that period. Based on these results of this research, I tried to outline what the mode of 'Sin-Pa' was, what it reflected, and what it desired for under the influence of the contemporary cultural conditions. I analysed double qualities of 'Sin-Pa' play as a dominant narrative and/or a resistant narrative considering its relationship with the people of colonized Korea. And also I examined the place of the 'Sin-Pa' play in the history of theatre and in the history of culture.
One of central issues in the Literature and Science discourses during the Victorian era is the relation of physiology to psychology. Many thinkers tackle the question of whether or not psychic phenomena can be reducible to their physiological bases. For instance, Victorian physiologist William Benjamin Carpenter claims that there should be a boundary between physiological and psychological qualities. Yet, his contemporary writer Grant Allen contends for the reduction of psychology into physiology. In the essay, I discuss Grant Allen's work Physiological Aesthetics (1877) so as to eventually problematize his physiological reductionism. I especially highlight the paradox of his physiological aesthetics. In order to clarify my argument, I introduce two concepts: evolutionary aesthetics and physiological reductionism. On the one hand, Allen argues for the development of aesthetic appreciation. The gradual evolution from gaudy to serene colors, for instance, reflects the fine differentiation of sensory organs. He believes that the existence of varied aesthetic pleasures corresponds to the evolution of sensory nerve structures. Nonetheless, Allen ironically gives more weight to the commonality of aesthetic experiences than to this teleological ordering of aesthetic experiences. He argues that there is no fundamental difference among humans in terms of their aesthetic assessments. Furthermore, there is even no essential distinction among plants, animals, and humans in light of their aesthetic appraisals, he states firmly. Although he asserts the gradual advance of aesthetic feelings caused by the intricacy of nervous systems, he simultaneously trivializes the evolution of aesthetic appraisal. In the essay, I highlight this paradox in Allen's physiological aesthetics. It should be underscored, lamentably enough, that Allen seeks biological purity by erasing fine lines among physiology, psychology, and sociality. He estranges aesthetic experiences from subjective variations and their socio-cultural contexts. He makes great efforts to eliminate individual differences and socio-cultural specificities in order to extremely biologize aesthetic experiences. Hence, Allen's physiological aesthetics is marked as the politics of physiological purification.
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