Secondary metabolism in actinomycetes has been known to be controlled by a small molecule, ${\gamma}$-butyrolactone autoregulator, the binding of which to each corresponding receptor leads to the regulation of the transcriptional expression of the secondary metabolites. We expected that expression of an autoregulator receptor or a pleiotropic regulator in a non-host was to be gained insight of effective production of new metabolic materials. In order to study the function of the receptor protein (seaR), which is isolated from Saccharopolyspora erythraea, we introduced the seaR gene to Streptomyces coelicolor A3(2) as host strains. An effective transformation procedure for S. coelicolor A3(2) was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a ${\varphi}C31$-derived integration vector, pSET152, which contained int, oriT, attP and $ermEp^*$ (erythromycin promotor). Therefore, the pEV615 was introduced into S. coelicolor A3(2) by conjugation and integrated at the attB locus in the chromosome of the recipients by the ${\varphi}C31$ integrase (int) function. Exconjugant of S. coelicolor A3(2) containing the seaR gene was confirmed by PCR and transcriptional expression of the seaR gene in the transformant was analyzed by RT-PCR. In case of S. coelicolor A3(2), a phenotype microarray was used to analyze the phenotype of transformant compared with wild type by seaR expression. After that, in order to confirm the accuracy of the results obtained from the phenotype microarray, an antimicrobial susceptibility test was carried out. This test indicated that sensitivity of the transformant was higher than wild type in tetracycline case. These results indicated that some biosynthesis genes or resistance genes for tetracycline biosynthesis in transformant might be repressed by seaR expression. Therefore, subsequent experiments, analysis of transcriptional pattern of genes for tetracycline production or resistance, are needed to confirm whether biosynthesis genes or resistance genes for tetracycline are repressed or not.
Proceedings of the Korean Society of Embryo Transfer Conference
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2007.05a
/
pp.39-50
/
2007
Sexing from bovine embryos fertilized in vitro implicates a possibility of the sex controlled cattle production. This study was carried out to produce the sex determined cattle through the embryonic sexing by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe constructed from the btDYZ-1 sequence. Using this probe, a male-specific signal was detected on 100% of Y-chromosome bearing metaphase specimens. The analyzable rate of embryonic sexing by FISH technique was about 93% (365/393) regardless of embryonic stages. As tested single blastomere by FISH and then karyotype with their biopsied embryos, the accuracy of sex determination with FISH was 97.6%. We tried the embryo transfer with sex determined embryos on 15 cattle. Among them, the 5 cattle delivered calf with expected sex last year.
Journal of the Korea Academia-Industrial cooperation Society
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v.5
no.2
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pp.141-149
/
2004
The aim of this study was to test the validity of the hypothesis that the karyotypes of four species of Lepilemuridae were formed spontaneously from their ancestral hybrid karyotype. Hypothetical ancestral haploid Karyotype of Lepilemuridae is composed of 18 autosomes and X chromosomes. Lepilemur mustelinus karyotype has four tandem fused chromosomes and one Robertsonian translocated chromosome pairs. Lepilemur septentrionalis septentrionalis karyotype has only two pairs of translocated chromosomes. We reconstruct and suggest ancestral karyotype of LMU(ancLMU) and LSS(ancLSS), from which all four studied species were derived. Hybrids of ancLMU and ancLSS were formed and produce differently fused equilibrated gametes via circular form arrangement during gametogenesis. Five unit of trivalent homologous chromosome pairs were engaged in a circular form to give new gamete corresponding to the karyotype of L. dorsalis, orientation of one unit of trivalent was inversed in the circle to gave new gamete corresponding to the karyotype of L. leucopus. Seven homologous chromosome pairs were engaged in circular form to give haploid karyotype of Lepilemur ruficaudatus. Only one homologous chromosome pair is dissociated and the other chromosome pairs rearranged in the circle to form haploid karyotype of Lepilemur edwardsi. The new gametes could be produced from these circular forms. When the new gamete fertilized with the same type of gamete, The new homozygote is produced as existing L. dorsalis, L. leucopus, L. edwardsi and L. ruficaudatus. These results support the theory that new species could be formed in hybrid population through activated chromosome fusion, chromosome rearrangement in circular from at zygotene stage and production of equilibrated gametes to form homozygote new species.
Purpose: Water is magnetically charged upon contact with a magnet. Although magnetic water products have been promoted since the 1930's, they have not received wide acceptance since their effectiveness is still in question; however, some have reported their therapeutic effects on the body, especially the digestive, nervous, and urinary systems. Methods: In this study, the effect of magnetized water on glycemic control of 14 diabetic mice (CB57BK/KsJ-db/db) in comparison with 10 control mice (CB57BK/KsJ-db/+(db/+)) was investigated. Seven diabetic control (DMC) mice and seven diabetic mice + magnetized water (DM+MW) were kept for 16 weeks, followed by intraperitoneal glucose tolerance test (IPGTT). Weekly blood glucose was measured from tail veins. Blood obtained from heart puncture was used for HbA1c analysis. Results: Blood glucose level showed a significant difference starting from the $10^{th}$ week of study ($496.1{\pm}10.2mg/dl$ in DMC vs. $437.9{\pm}76.9mg/dl$ in DM+MW). Blood glucose followed by IPGTT showed no significant difference between groups at 0, 30, 60, 90, and 120 min, although glucose level at 180 min was significantly reduced in DM+MW mice. Plasma insulin level in DM+MW groups was only 39.5% of that of DMC groups ($5.97{\pm}1.69ng/ml$ in DMC vs. $2.36{\pm}0.94ng/ml$ in DM+MW). Levels of HbA1c were 12.4% and 9.7% in DMC and DM+MW groups, respectively. Conclusion: These results show the promising therapeutic effect of magnetized water in regulating blood glucose homeostasis; however, long-term supplementation or mechanistic study is necessary.
For the head and neck radiotherapy, the technique of half beam using independent collimator is very useful to avoid overlapping of fields particularly when the lateral neck fields are placed adjacent to anterior supraclavicular field. Also abutting photon field with electron field is frequently used for the irradiation of posterior neck when tolerable dose on spinal cord has been reached. Using 6 MV X-ray and 9 MeV electron beams of Clinac1800(Varian, USA) linear accelerator, we performed film dosimetry by the X-OMAT V film of Kodak in solid water phantom and the dose distribution at beam center of 2 half beams further examined according to depths(0 cm, 1.5 cm, 3 cm, 5 cm) for single anterior half beam and anterior/posterior half beam. The dose distribution to the junction line between photon and electron fields was also measured. For the single anterior half beam, the absorption doses at 0.3 cm, 0.5 cm and 1 cm distances from beam center were 88%, 93% and 95% of open beam, respectively. In the anterior/posterior half beams, the absorption doses at 0.3 cm, 0.5 cm and 1 cm distances from beam center were 92%, 93% and 95% of open beam, respectively At the junction line between photon and electron fields, hot spot was developed on the side of the photon field and a cold spot was developed on that of the electron field. The hot spot in the photon side was developed at depth 1.5 cm with 7 mm width. The maximum dose of hot spot was increased to 6% of reference doses in the photon field. The cold spot in the electron side was developed at all measured depths(0.5 cm-3 cm) with 1-12.5 mm widths. The decreased dose in the cold spot was 4.5-30% of reference dose in the electron field. With above results, we concluded that when using electron beam or independent jaw for head and neck radiotherapy, the hot and cold dose area should be considered as critical point.
Objective: To evaluate the significance and efficacy of trans-cervical fallopian tube catheterization (TFTC) in diagnosis and optimal treatment modality for tubal blockage. Methods: The retrospective study was performed in those underwent TFTC from January 2005 to December 2009. A total of 342 fallopian tubes in 215 patients which showed tubal blockage in hysterosalpingography (HSG), were subjected to TFTC. Recanalization rate (RR) was compared according to portion of tubal blockage; proximal, isthmic and distal portion and blockage type; tapering, concave, and convex type. Results: In total, RR was 72.5% (248/342 tube). According to the portion of tubal blockage, RR was 83.8% in proximal, 45.6% in isthmic and 100% in distal portion. RR was 92.3% in tapering, 80.2% in concave and 25.5% in convex type, respectively. There were 98 pregnancies in 156 patients after successful recanalization, which shows 62.7% pregnancy rate. Conclusio: TFTC were capable of recanalizing tubal blockage in 248 of 342 tubes in 156 of 215 patients (72.5%). The RR was increased with proximal portion and tapering type tubal blockage.
The Sobeaksan Gneiss Complex in the Punggi area is composed of mainly mignatitic gneiss, porphyroblastic gneiss, garnet granitic gneiss and biotitie granitic gneiss. Metamorphic grade increase gradually from the amphibolite facies of northwestern part to the granulite facies of southwestern part in the study area. Representative mineral assemblage in the amphibolite facies is biotite-muscovite-K-feldspar-plagioclase$\pm$garnet$\pm$epidote, needle shape or fibrous sillimanite occur in transitional zone from the amphibolite facies to the granulite facies. In the granulite facies, the garnet-Opx granulite shows garnet-orthopyroxene-biotite-plagioclase, the metabasite shows clinopyroxene-plagioclase$\pm$hornblende$\pm$orthopyroxene$\pm$garnet and the migmatitic gneiss shows garnet-biotite-sillimanite-cordierite$\pm$spinel as representative mineral assemblage. Retrograde metamorphism after the granulite facies metamorphism made corindum and andalusite in the migmatitic gneiss and the thin layer garnet between clinopyroxene and plagioclase in the metabasites. The peak P-T conditions of the migmatitic gneiss and the garnet-Opx granulite are $916^{\circ}C$/6.6 kb and $826^{\circ}C$/6.3 kb, respectively. The P-T condition of biotite and plagioclase inclusion, which indicates the progressive condition of the granulie facies, within garnet is $866^{\circ}C$/7.5 kb and that of rim composition of garnet and biotite is $726^{\circ}C$/4.6 kb, which infer the clockwise P-T path of the granulite facies metamorphism. The temperatures caculated by the rim composition of garnet and biotite in the migmatitic gneiss and garnet granitic gneiss have a wide range of $556-741^{\circ}C$, which indicate that the retrograde metamorphism after the granulite facies metamorphism has effected differently. It is difficult to determine the P-T condition of the biotite granitic gneiss because less occurrence and higher spessartine content of garnet. The P-T condition of the thin layered garnet between clinopytoxene and plagioclase in the metabasite is $635-707^{\circ}C$/4.1-5.3 kb. This texture indicates the isobaric cooling(IBC) condition of the retrogressive metamorphism. As a result, the metamorphic evolution of the Punggi area has undergone the isobaric cooling after the granulite facies metamorphism which has undergone the clockwise P-T path.
Kim, Jong-choon;Lee, Hyun-sook;Yun, Hyo-in;Chung, Moon-koo
Korean Journal of Veterinary Research
/
v.40
no.2
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pp.361-371
/
2000
It has been recently reported that 2-bromopropane (2-BP) induces male reproductive toxicity in both human and experimental animals. However, delayed effects of 2-BP on male reproductive system have not been investigated in detail. The present study was conducted to investigate the testicular toxicity of 2-BP and to determine the recovery of normal spermatogenesis in Sprague-Dawley rats. Male rats aged 5 weeks were administered 1,000mg/kg 2-BP by gavage daily for 4 weeks and sacrificed sequentially at 1, 2, 3, 4 and 12 weeks after initiation of 2-BP treatment. Testicular toxicity was evaluated qualitatively by histopathological examinations and quantitatively by reproductive organ weights, spermatid head count, and repopulation index. In the 2-BP treated rats, the body weights was significantly suppressed and the weights of testes and epididymides were also decreased in a time-dependent manner. On histopathological examination, spermatogonia in stages I-VI and preleptotene and leptotene spermatocytes in stages VII-IX were strongly depleted at 1 week of dosing. Spermatogonia were depleted extensively in all spermatogenic stages at 2 weeks. Continuing with the evolution of spermatogenic cycle, zygotene spermatocytes, pachytene spermatocytes, and round spermatids were sequentially depleted at 2, 3, and 4 weeks of dosing due to the depletion of their precursor cells. Vacuolization of Sertoli cells and spermatid retention were also observed at all time points, suggesting that 2-BP induced Sertoli cell dysfunction. At 12 weeks, after 8 weeks recovery, most of the tubules appeared severely atrophic and were lined by Sertoli cells only. Leydig cell hyperplasia in the interstitial tissue was also found. In addition, dramatic reductions in the number of spermatid heads and repopulation index were observed, indicating that 2-BP-induced testicular injury is irreversible. These results indicate that 4 weeks repeated-dose of 1,000mg/kg 2-BP results in a progressive germ cell loss due to the depletion of spermatogonia followed by long-term testicular atrophy in SD rats.
To understand the relationship between coat color inheritance patterns and genotypes of Extension (E) and Agouti (A) loci in cattle, the genotypes for melanocortin-1 receptor (MC1R) and agouti signaling protein (ASIP) were analyzed in Hanwoo, Jeju black cattle (JBC), and their crossbred progeny. Three MC1R alleles ($E^D$, $E^+$, and e) were found in the black-colored JBC population. JBC had no recessive homozygotes (e/e), but this genotype was predominant in the Hanwoo breed. However, MC1R $E^+$/e Hanwoo did not produce a black coat color as they appeared either as brown or solid red. For ASIP, three genotypes (A/A, A/$A^{Br}$, and $A^{Br}/A^{Br}$) were determined by insertion/deletion of an L1-BT element in Hanwoo. The ASIP $A^{Br}$ allele was rarely observed, and no ASIP $A^{Br}/A^{Br}$ homozygotes were detected in the JBC population. Cattle carrying ASIP $A^{Br}$ did not show any agouti-like brindle pigmentation patterns in either breed or their progeny. The coat colors of the crossbred progeny were discriminated by two colors, yellowish-brown versus dark-brown or black, and their coat colors were directly related to the genotypes of the Extension locus, yellowish-brown (e/e) and dark-brown or black ($E^+$/e), but not to the Agouti locus. ASIP genotypes probably did not affect coat color development in the Hanwoo or crossbred progeny. Our results suggest that the ASIP genotypes do not play key roles in coat color variation, but the MC1R genotypes do direct the phenotypes of Hanwoo, JBC, and their progeny.
The level of genetic variation and relationships in three native Korean goat populations (Dangjin, Jangsu, and Tongyeong) as well as the populations of a farm were analyzed, based on 30 microsatellite markers. A total of 277 distinct alleles were observed across the four goat populations, and 102 (36.8%) of these alleles were unique to only one population. The mean observed heterozygosity and polymorphism information content were calculated as 0.461~0.651 and 0.462~0.679, respectively. In the NJ tree constructed based on Nei's $D_A$ genetic distance, the four populations represented four distinct groups. However, the genetic distances between each Korean native goat population and the farm population were two times those among the three native Korean breeds. The genetic structure within the three Korean native goat populations was also investigated. Cluster analysis, using the STRUCTURE software, suggested three clusters. The molecular information of genetic diversity and relationships in this study will be useful for the evaluation, conservation, and utilization of native Korean goat breeds as genetic resources.
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