Purpose : X-linked agammaglobulinemia(XLA) is an immunodeficiency caused by abnormalities in Bruton's tyrosine kinase(Btk), and is characterized by a deficiency of peripheral blood B cells. We studied cytoplasmic expression of Btk protein and analyzed the Btk gene in peripheral blood mononuclear cells(PBMC) from three XLA families in Korea. Methods : Heparinized venous blood samples were collected from four XLA patients and additional family members in three unrelated XLA families. Mononuclear cells were separated from their blood and the intracellular Btk protein was characterized by a flow cytometry. The mutation analysis was performed using direct sequencing. Results : Cytoplasmic expression of Btk protein in monocytes was not detected in the patients with XLA. We observed a novel deletion and two point mutations within introns(intron 1 and intron 18) resulting in alternative splicings. In XLA family 2, a 980 bp deletion(from intron 9+191 T to intron 10-215 C) including exon 10 was found in patient P2. He was the only sporadic case in this study, because his mother and brother showed a normal Btk expression by flow cytometry. Conclusion : These identified genetic alterations support the molecular heterogeneity of Btk gene in XLA disease. Additionally, by means of flow cytometric analysis, we diagnosed three hypogammaglobulinemia patients as XLA. Advancements in diagnostic methods has facilitated a prompt and definite diagnosis of this disease.
Kang, Ho Bum;Ryoo, Seung Heui;Lee, Sang Hoon;Jeon, Byung Soon;Sang, Byung Chan
Korean Journal of Agricultural Science
/
v.25
no.1
/
pp.79-88
/
1998
This study was performed in order to apply to effective breeding of Korean native cattle on the molecular genetic level obtained from PCR and nucleotide sequencing analysis of BoLA DRB3 exon2 that has important roles in host immune defence. Genomic DNA used in this study was prepared from the blood of Korean native cattle in Korean Native Cattle Improvement Center of National Livestock Cooperation. The results obtained from this study are summarized as follows: 1. Genomic DNA extracted from the blood of Korean native cattle was subjected to electrophoresis on 1.5% agarose gel. Major band was bigger than 12.2kb, indicating that genomic DNA was well prepared for PCR. Amplified products of 284bp fragments was obtained the amplification of BoLA DRB3 exon2 gene by PCR. 2. Cloning of BoLA DRB3 exon2 of Korean native cattle with pCR2.1 vector was conformed by 300bp fragment from recombinent plasmid that restricted with enzyme digestion of EcoRI. 3. Homology of BoLA DRB3 exon2 alleles of parent was 82.0% between sire's alleles and 90.1% between dam's alleles. 4. In pedigree analysis using BoLA DRB3 exon2 gene, sequencing result of BoLA DRB3 exon2 genes showed inheritance by Mendelian mode through the parents to their offspring. 5. Taking together those experimental results, pedigree was confirmed on the basis of sequencing for the alleles of parents and offspring. This knowledge by the molecular biological approach could be served for the improvement of Korean native cattle.
A new commercial strain "Mongdol" of oyster mushroom was developed by hyphal anastomosis. It was improved with hybridization between monokaryotic strain derived from Pleurotus ostreatus ASI 0627 and dikaryotic strain derived from P. ostreatus ASI 2929. The optimum temperature of mycelial growth and fruiting body development were $25{\sim}30^{\circ}C$ and $12{\sim}18^{\circ}C$, respectively. When two different media including PDA (potato dextrose agar medium) and MCM (mushroom complete medium) were compared, the mycelial growth of this mushroom was faster in MCM than in PDA. Similar result was observed with the control strain P. ostreatus ASI 2504. Analysis of the genetic characteristics of the new cultivar "Mongdol" showed a different DNA profile as that of the control strain ASI 2504, when RAPD (Random Amplified Polymorphic DNA) primer URP3 and URP6 were used. Fruiting body production per bottle was about 106 g using demonstration farms. The color of pileus was blackish gray and the stipe was long. Therefore, we expect that this new strain "Mongdol" will satisfy the consumer's demand for high quality mushrooms.
Oyster mushroom is a type of mushroom that is commonly cultivated and consumed in Korea. P. ostreatus 'Suhan' is a preferred cultivar for many mushroom farmers because it has a dark pileus and thick stipe. However, as it is very sensitive to environmental conditions, farmers consistently demand an alternative cultivar. To develop a new cultivar, the parental strains KMCC01680 ('Suhan') and KMCC00478 ('Gosol') were selected from various collected P. ostreatus strains by cultivating genetic resources. P. ostreatus 'Heuksol' was developed by the method of Mon-Mon crossing between monokaryotic strains derived from 'Suhan' and 'Gosol'. Thirty strains of 174 crossed strains were initially selected by cultivation experiments. After bulk cultivation tests, 'Heuksol' was selected. The nuclear DNA profile of 'Heuksol' was similar to those of the parental strains, 'Suhan' and 'Gosol', when RAPD (random amplified polymorphic DNA) primers and UPF (Universal PCR Fingerprinting) 2, 3, and 4 were used. The optimum temperature for mycelial growth was $30^{\circ}C$ for 'Heuksol', but medium-high temperatures were also appropriate, especially $13-20^{\circ}C$. The fruiting body production per bottle (1,100 mL) was approximately 140.8 g. When compared to the control strain 'Suhan', the thickness of the stipe of 'Heuksol' was greater than that of 'Suhan' (13.5 mm vs 9.4 mm). The pileus diameter of 'Heuksol' was similar to that of 'Suhan' and the pileus thickness of 'Heuksol' and 'Suhan' was 19.7 mm and 21.8 mm, respectively. 'Heuksol' had more a productive stipe number than 'Suhan' and the pileus of 'Heuksol' was dark gray, even at high temperatures. Therefore, it was suggested that this new cultivar, 'Heuksol', could provide an alternative to 'Suhan' and contribute to the profit of oyster mushroom farms.
Lysine is the first limiting essential amino acid in cereals for humans and monogastric animals, although its content is generally low. A chemically induced high-lysine barley mutant, M98, has an agronomically undesirable shrunken endosperm trait. In order to obtain detailed insight into the atypical traits of M98 grains, we characterized amino acid composition and protein profiles of M98 and its parent cultivar Chalssalbori. Among a total of 16 amino acids, the percentage of each of the 7 amino acids, including lysine, was 1.2~1.8 times higher in M98, comparing to Chalssalbori. The percentage of proline and its precursor, glutamic acid, in M98 was about the half of that of the amino acids in Chalssalbori, but arginine synthesized from glutamic acid was 1.8 times higher in M98, compared that in the parent cultivar. Theses results indicated that the mutation in M98 grains might alter the proportion of amino acids linked to each other in a biosynthetic pathway. A comparison of grain proteome profiles between Chalssalbori and M98 revealed 70 differentially expressed protein spots, where 45 protein spots were up-regulated and 25 protein spots down-regulated in M98 compared to those in Chalssalbori. Of these changed protein spots, 53 were identified using nano-electrospray ionization liquid chromatography mass spectrometry. Most of these identified proteins were involved in various biological processes. In particular, 28 protein spots such as ${\beta}$-amylase, serpins and B3-hordein were identified as proteins associated with the atypical traits of M98. It was thought that a genetic study on the unique protein profile of M98 would be needed to develop an agronomically feasible barley cultivar with high-lysine trait.
The two most common mushroom species grown in Korea are pearl oyster mushroom (Pleurotus ostreatus) and king oyster mushroom (P. eryngii). In recent years, the production of king oyster mushroom greatly increased due to the automation of the cultivation facilities, and it became a major export mushroom owing to its excellent shelf life. However, the increase in the production of king oyster mushroom led to a decline in its market price; thus, necessitating the development of new mushroom species that could replace king oyster mushroom, to diversify the mushroom market for the benefit of both, the producers and the consumers. The Mushroom division at the National Institute of Horticultural & Herbal Science (NIHHS) reported the development of a new interspecific hybrid between P. ferulae and P. tuoliensis, referred to as 'Creamy.' Two parental strains KMCC00430 (Bisan2ho, P. ferulae) and KMCC00461 (P. tuoliensis) were selected based on the results of genetic resource analysis, and their monokaryons were collected. About 1,000 Mon-Mon crosses were performed and 73 of them were selected. Following repeated cultivation tests and strain analyses, we selected strain 7773, which had a bright creamy pileus and a thick straight stipe, and named it 'Creamy.' Optimum temperature for mycelial growth of Creamy was 25-30℃, and that for fruiting body growth was 16℃. The pileus, which had a brighter creamy color, was small in size with a diameter of 61.2 mm. Although it was cultivated in suboptimal conditions, such as low temperature and high CO2 concentration, Creamy was characterized by its straight and smooth stipe. Field production tests and further analyses indicated that the yield of Creamy was 5% higher than that of Baekhwang. It is expected that Creamy, the new interspecific hybrid with a bright creamy pileus and a pleasant flavor, will help create new opportunities for mushroom farmers and diversify the mushroom market.
Bentonites from the Janggi Group of the Lower Miocene age from the Geumgwangdong area, Korea, have been studied for mineralogical and genetic characterization. The Janggi Group is subdivided, in ascending order, into the Janggi Conglomerate, the Nuldaeri Tuff, the Geumgwangdong Shale, the Lower Coal-bearing Formation, the Basaltic Tuff, and the Upper Coalbearing Formation. Bentonites occur as thin or thick beds in all sedimentary units of the Janggi Group, except for the Janggi Conglomerate. Significant bentonite deposits are found in the Nuldaeri Tuff, the Lower Coal-bearing Formation and the Basaltic Tuff. Bentonites consist mainly of smectite (mainly montmorillonite), with minor quartz, cristobalite, opal-CT and feldspar. Occasionally, kaolinite, clinoptilolite or gypsum is associated with bentonites. Bentonites were studied by the methods of petrographic microscopy, X-ray diffraction, thermal analysis (DT A and TG), infrared absorption spectroscopic analysis, SEM, intercalation reaction, and chemical analysis. Smectites commonly occur as irregular boxwork-like masses with characteristic curled thin edges, but occasionally as smoothly curved to nearly flat thin flakes. Most of smectites have layer charge of 0.25-0.42, indicating typical montmorillonite. Crystal-chemical relations suggest that Fe is the dominant substituent for Al in the octahedral layer and there are generally no significant substituents for Si in the tetrahedral layer. Ca is the dominant interlayer cation in montmorillonite. Therefore, montmorillonite from the study area is dioctahedral Ca-montmorillonite. Occurrence and fabrics of bentonites suggest that smectites as well as cristobalite, opal-CT and zeolites have been formed diagenetically from tuffaceous materials. The precursor of smectites is trachytic or basaltic tuff. Smectites derived from the former contain relatively more $Al_2O$ a and less $Fe_2O_3$ than those from the latter.
Ferritin is known to be the principle iron-storage protein in a wide variety of rganisms. The electrophoretic mobility and immunological cross-reactivity of dog splenic ferritin were compared with those of horse, bovine, and pig splenic ferritin after isolation using heat treatment, salting out, column chromatography, and ultrafiltration. These isolation methods allowed the recovery of $\~84{\mu}g$ of the ferritin per g of spleen. The iron content in the dog ferritin was $22.7\%$, which appeared to be higher than those in the other mammalian ferritins tested. The electrophoretic mobility of the dog ferritin under nondenaturing conditions was similar to its bovine counterpart, whereas it was more identical to pig and horse ferritins on an SDS-polyacrylamide gel. The molecular weight of the dog ferritin subunit was 19.5 kDa on an SDS-polyacrylarnide gel, and the subunit was unable to bind with iron. The polyclonal anti-dog ferritin raised in rats was able to cross-react with the pig, bovine, and horse ferritins, upon Ouchterlony double immunodiffusiion. A Western blot analysis also revealed that the anti-dog ferritin, which specifically bound with the dog ferritin subunit, could also recognize the horse, bovine, and pig ferritin subunits and the maximum cross-reactivity was exhibited with the pig ferritin subunit, indicating that the dog ferritin is immunochemically more similar to the pig ferritin than its other mammalian counterparts. Accordingly, these results elucidate the biochemical and immunochemical characteristics of dog ferritin that might have a potential to be applied as an oral iron supplement to treat iron deficiency anemia.
Kim, Ki-Yong;Choi, Young-Jin;Rim, Yong-Woo;Seong, Byung-Ryul;Lee, Sang-Jin;Yang, Joo-Sung;Hahn, Bum-Soo;Kim, Jong-Bum;Lee, Byung-Hyun
Journal of The Korean Society of Grassland and Forage Science
/
v.24
no.4
/
pp.341-346
/
2004
To determine lethal temperature of birdsfoot trefoil(BFT) and italian ryegrass(IRG) at heat-stressed conditions, seedlings grown in a small pots fur 4 weeks were subjected to different temperature regimes of heat treatment. No apparent damage was observed BFT and IRG were treated at 45, 50 or $60^{\circ}C$ for 1 h. And also heat treatments at 60, 65 and $70^{\circ}C$ for 1 h, both of them were withered and showed damage symptom on their leaves but it was not lethal conditions for the whole plants. By contrast, most of plants were prominently withered within one day after heat treatment at $80^{\circ}C/60min$. When BFT was exposed to $80^{\circ}C/60$ min, they were died within 6 days but there was found that new shoots were regenerated from the plants that had been treated at $80^{\circ}C$ within 55 min. IRC was also died within 2 days that exposed to $80^{\circ}C/20$ min but there was found that new shoots were regenerated from the plants that had been treated at $80^{\circ}C$ within 15 min. These results indicate that heat killing temperatures of BFT and IRG plants are $80^{\circ}C/60$ min an $80^{\circ}C/20$ min respectively. Simple viability assay system established in this study will be useful for selection and characterization of heat-tolerant transgenic BFT and IRG plants.
Yoo, Young Bok;Lee, Sang Cheol;Kim, Eun Jung;Kong, Won Sik;Jang, Kab Yeul;Shin, Pyung Gyun
Journal of Mushroom
/
v.7
no.3
/
pp.130-134
/
2009
To develop new white variety of Oyster mushroom, all white varieties which have been collected and kept in the lab were revived and screened their cultural characteristics. 84 intra-specific Oyster mushroom hybrids between the white-colored mutants Suhan and Wonhyeong were developed using hyphal anastomosis technique in 2007. The Po2007-63 ($2842-7{\times}0205-7$) was shown the best cultural characteristics, selected to be a new variety and named as 'Goni'. The new commercial strain, 'Goni' has white pilei and grows well under spring and autumn conditions in Korea. The fruiting bodies of 'Goni' are of an excellent quality in that not only the stipe is thick and long but also the pileus is small and hard. The optimum temperatures for mycelial growth and fruiting body development were $25-30^{\circ}C$ and $10-16^{\circ}C$, respectively. Time period required for the initiation of the first fruiting body is about 3 to 5 days depending on the temperatures. The shape of fruiting body is thin funnel shape. Fruiting body production per bottle was about $91{\pm}13$ g which is almost 97% quantity compared to that of other variety 'Miso'. Relatively low temperature incubation ($11^{\circ}C$) resulted in the development of better quality of 'Goni' mushrooms. When two different media including potato dextrose medium and mushroom complete medium were compared, the growth of mushroom were much faster in mushroom complete medium at $20-25^{\circ}C$, but not at $25^{\circ}C$. Similar results were observed with other variety 'Miso'. Analysis of the genetic characteristics of the new commercial strain 'Goni' showed a major DNA profile as that of the parental Suhan when primer URP 1 was used, but different to 'Miso' that was used as a control. When screens were performed with primer URP 2, DNA patterns were similar both to that of the parents and 'Miso'. This new variety of the white Oyster mushroom has a clean and fresh image that corresponds well to "health food". We therefore expect that this new strain will satisfy the consumers demand for variety and excellent mushrooms.
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