• Journal of Plant Biotechnology
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• v.30 no.1
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• pp.13-18
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• 2003

This study was carried out to investigate the expression patterns of foreign gene that controlled by tuber-specific patatin promoter in transgenic potatoes. Potato leaf disc cultured in vitro were transformed by the Agrobacterium strain LBA4404 containing pBl121 or pATGUS from potato cv. Desiree. In order to select the transgenic lines, gene-specific primers deduced from the NPTII were synthesized and used for polymerase chain reaction. The down part of the putative transgenic potatoes was transplanted weekly onto sucrose-enriched medium to accelerate the microtuber formation. RNA gel blot analysis was performed on the total RNAs obtained from tuber that had been harvested at a week interval. Also, histochemical assay was observed in the explants transformed with either pBI121 or pATGUS. Results showed that the transgenic plant containing pATGUS expressed GUS transcripts mainly at the tuber, not in stem, with the highest expression level in 5 weeks-grown microtubers. In contrast to pATGUS plants, the transformed plants with pBI121 showed an equal expression pattern throughout the whole developing stages. Consistent with RNA gel blot analysis, histochemical GUS staining and enzyme activity exhibited pATGUS transcripts were at the highest level in 5 weeks cultures. From these results, we suggest that the best stage to analyze the foreign gene introduced by patatin promoter into potato plants is at 5 weeks cultures after tuber formation.

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.163-169
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• 2001

OSBA(oocytes-sperm binding assay) is a tool developed for rapid test of optimal condition of IVF medium and protein source by binding ability of mouse sperm and egg. Mouse oocyte-cumulus complexes were prepared by removing of the cumulus cells with 0.1% hyaluronidase. 10$\pm$2 oocytes per 30 ${mu}ell$ medium drop were inseminated with 3 ${mu}ell$ sperm suspension and were cultured f3r 3 hours and 24 hours, respectively. And the oocytes were recovered gently and the No. of sperm bound on oocytes were counted. In the Exp. 1, the ratio of oocytes bound with one sperm at least were 60.2%(50/83), 2%(2/77) and 100%(79/79) in the medium with no protein, FBS(15%, v/v) and BSA(0.4%. w/v), respectively, Fetal bovine serum(FBS) seriously inhibited sperm binding on oocyte, although bovine serum albumin(BSA) promoted the binding ability. The inhibiting effect of FBS was dependent on the concentration of FBS. The sperm binding ability according to oocyte maturity was tested in the Exp. 2. There was no significant difference between Met. II (mature) and Met. I (intermediate mature) oocytes in the number of oocytes bound with sperm and the number of sperm bound on oocytes. Finally, in Exp. 3, two batches of Ham's F10 medium with good and poor quality by OSBA were tested (The ratios of embryos developed from PN 1-cell stage to hatched blastocyst; 25% vs. 70%). In the medium with good quality, sperm binding ability was significantly increased (P ＜ 0.05). The ratio of oocytes bound with one sperm at least was 66% and 90% in the medium with poor and good quality, respectively. Conclusively, It was possible to test IVF medium condition rapidly and easily by OSBA.

• Journal of fish pathology
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• v.18 no.1
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• pp.67-80
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• 2005

Phosphoinositide-specific phospholipase $C\delta$ $PLC\delta$) plays an important role in many cellular responses and is involved in the production of second messenger. The present study was conducted to obtain the biochemical characteristics of the expressed recombinant $PLC\delta$ in E. coli cloned from Misgurnus mizolepis and partially purified $PLC\delta$ enzymes from liver tissues of M. mizolepis (wild ML-$PLC\delta$). The ML $PLC\delta$ gene was cloned and expressed under the previous report (Kim et al., 2004), and purified the recombinant protein by successive chromatography using $Ni^{2+}$-NTA affinity column and gel iltration FPLC column. The wild ML-$PLC\delta$ protein was solublized with 2 M KCI and purified by successive chromatography on open heparin-Sephagel and analytical TSKgel heparin-5PW. Both the recombinant and wild ML-$PLC\delta$ form of protein showed a concentration-dependent PLC activity to phosphatidylinositol 4,5-bis-phosphate (PIP$_2$) or phosphatidylinositol (PI). Its activity was absolutely $Ca^{2+}$- dependant, which was similar to mammalian $PLC\delta$ isozymes. Maximal PI-hydrolytic activations of recombinant and wild ML- TEX>$PLC\delta$ was at pH 7.0 and pH 7.5, respectively. In addition, the enzymatic activities of recombinant and wild ML-$PLC\delta$ were increased in concentration-dependent manner by detergent, such as sodium deoxycholate SDC), phosphatidylethanolamine (PE) and phosphatidylcholine (PC). The activities decreased in contrast by a polyamine, such as spermine. Western blotting showed that several types of $PLC\delta$ isozymes exist in various organs. Taken together our results, it suggested that the biochemical characteristics of ML-$PLC\delta$ are similar with those of mammalian $PLC\delta1$ and ${\delta}3$ isozymes.

• Journal of Life Science
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• v.32 no.2
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• pp.135-141
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• 2022

Thymosin β-4 (TB4) is a small peptide composed of 43 amino acids. To obtain sufficient biologically active mouse TB4 economically, we cloned and overexpressed this gene in an Escherichia coli system. With the isopropyl β-D-1-thiogalactopyranoside induction of the E. coli transformant, TB4 fusion protein with intein- and chitin-binding domain was successfully expressed in the soluble fraction within the E. coli cell. The TB4-intein - chitin-binding domain fusion protein was purified from the soluble fraction of E. coli cell lysate. The affinity chromatography with chitin beads and dithiothreitol-mediated intein self-cleavage reaction releases the TB4 peptide into the stripping solution. Sodium dodecyl sulphate - polyacrylamide gel electrophoresis and Western blot analyses were used to confirm that the recombinant TB4 peptide was produced with the expected size of 5 kDa. We found that the recombinant TB4 stimulated cell migration in the transwell plate chamber assay. After 18 hr of the treatment of the recombinant TB4 with 1 ng/ml concentration, the migration of the HT1080 cell was increased by 20% compared with that of the chemically synthesized TB4. The recombinant TB4 was also observed to promote the healing of a wound area in C57BL/6 mice by as high as 35% compared with that of the chemically synthesized TB4. These results suggest that the recombinant TB4 has better biological activity for cell migration and wound healing than that of the chemically synthesized TB4 peptide.

• The Journal of Korean Physical Therapy
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• v.10 no.1
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• pp.7-21
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• 1998

An analysis of 101 stroke patients who were enrolled in 10 hospitals of Cities of Pusan, Taegu, and Andong from November 1, 1996 to April 31. 3997 was conducted using the modified Barthel Index(BI) and the adapted PULSES profile index(PS) to evaluate their function. Patients were examined at the following intervals: Initial assessment, one month after initial. at discharge, and one month after discharge. The mean BI score of patients initial assessment was 27.18, and that of PS was 17.54. There were statistically significant between initial score and one month after initial (21.39: p<0.001), at discharge(37.47: p<0.001), one month after discharge(46.49: p<0.001). PS scores were also improved .significantly(-2.62, -4.52. and -6.26(p<0.001). And the score between dischange and one month after discharge was significant (9.01: p<0.001) and in PS score(-1.73: p<0.001). Age and BE score were significantly associated with the improved in BI score between initial and discharge(T3-T1)(p<0.05). Below age forty the Bower initial BI score showed significantly higher improvement(T3-T1) after physical therapy(p<0.05). Initial Bl score, patients' attitude for physical therapy after discharge, age, and surgical operation were significantly associated with the improvement of BI score between initial and one month after discharge(T4-T1)(p<0.05). The lower initial BI score, patients' positive attitude for physical therapy after discharge, below age forty. and no surgical operation showed significantly higher improvement(p<0.05). BI score at discharge, side of hemiparesis and religion were significantly associated with the improvement of BI score between at discharge and one month after discharge(T4-T3) (p<0.05). The lower BI score at discharge. left aide of hemiparesis, with religion showed significantly higher improvement (p<0.05). Age, initial PS score were significantly associated with the improved in PS score between initial and discharge(T3-T1)(p<0.05). The higher initial PS score and below age forty showed significantly higher improvement(T3-T1)(p<0.05). Initial PS score, patient' attitude for physical therapy after discharge, age, educational level, physical therapy hour after discharge, and surgical operation were significantly associated with the improvement of PS score between initial and one month after discharge(T4-T1)(p<0.05). The higher initial PS score, patients' positive attitude for physical therapy after discharge, below age forty, higher education, the shorter physical therapy hour, and no surgical operation showed significantly higher improvement(T4-T1)(p<0.05). PS score at discharge, educational level, patient' attitude far physical therapy after discharge, physical therapy hour after discharge, and gender were significantly associated with the improvement of PS score between discharge and une month after discharge(T4-T3) (p<0.05), The higher PS non at discharge, higher education, patients' positive attitude for Physical therapy after discharge, the shorter physical therapy hour, and male showed significantly higher improvement (T4-T3)(p<0.05). In conclusion, initial BI score and age were significantly associated with BI score improvement and initial PS score, age, and educational level were also significantly associated with PS score improvement in stoke patients.

• Magazine of the Korean Society of Agricultural Engineers
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• v.16 no.2
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• pp.3361-3394
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• 1974

The purpose of this thesis is to disclose some characteristics of water consumption in relation to the quantities of dry matters through the growing period for two statured varieties of paddy rice which are a tall statured variety and a short one, including the water consumption during seedling period, and to find out the various coefficients of evapotranspiration that are applicable for the water use of an expected yield of the two varieties. PAL-TAL, a tall statured variety, and TONG-lL, a short statured variety were chosen for this investigation. Experiments were performed in two consecutive periods, a seedling period and a paddy field period, In the investigation of seedling period, rectangular galvanized iron evapotranspirometers (91cm${\times}$85cm${\times}$65cm) were set up in a way of two levels (PAL-TAL and TONG-lL varieties) with two replications. A standard fertilization method was applied to all plots. In the experiment of paddy field period, evapotanspiration and evaporation were measured separately. For PAL-TAL variety, the evapotranspiration measurements of 43 plots of rectangular galvanized iron evapotranspirometer (91cm${\times}$85cm${\times}$65cm) and the evaporation measurements of 25 plots of rectangular galvanized iron evaporimeter (91cm${\times}$85cm${\times}$15cm) have been taken for seven years (1966 through 1972), and for TONG-IL variety, the evapotranspiration measurements of 19 plots and the evaporation measurements of 12 plots have been collected for two years (1971 through 1972) with five different fertilization levels. The results obtained from this investigation are summarized as follows: 1. Seedling period 1) The pan evaporation and evapotranspiration during seedling period were proved to have a highly significant correlation to solar radiation, sun shine hours and relative humidity. But they had no significant correlation to average temperature, wind velocity and atmospheric pressure, and were appeared to be negatively correlative to average temperature and wind velocity, and positively correlative to the atmospheric pressure, in a certain period. There was the highest significant correlation between the evapotranspiration and the pan evaporation, beyond all other meteorological factors considered. 2) The evapotranpiration and its coefficient for PAL-TAL variety were 194.5mm and 0.94∼1.21(1.05 in average) respectively, while those for TONG-lL variety were 182.8mm and 0.90∼1.10(0.99 in average) respectively. This indicates that the evapotranspiration for TONG-IL variety was 6.2% less than that for PAL-TAL variety during a seedling period. 3) The evapotranspiration ratio (the ratio of the evapotranspiration to the weight of dry matters) during the seedling period was 599 in average for PAL-TAL variety and 643 for TONG-IL variety. Therefore the ratio for TONG-IL was larger by 44 than that for PAL-TAL variety. 4) The K-values of Blaney and Criddle formula for PAL-TAL variety were 0.78∼1.06 (0.92 in average) and for TONG-lL variety 0.75∼0.97 (0.86 in average). 5) The evapotranspiration coefficient and the K-value of B1aney and Criddle formular for both PAL-TAL and TONG-lL varieties showed a tendency to be increasing, but the evapotranspiration ratio decreasing, with the increase in the weight of dry matters. 2. Paddy field period 1) Correlation between the pan evaporation and the meteorological factors and that between the evapotranspiration and the meteorological factors during paddy field period were almost same as that in case of the seedling period (Ref. to table IV-4 and table IV-5). 2) The plant height, in the same level of the weight of dry matters, for PAL-TAL variety was much larger than that for TONG-IL variety, and also the number of tillers per hill for PAL-TAL variety showed a trend to be larger than that for TONG-IL variety from about 40 days after transplanting. 3) Although there was a tendency that peak of leaf-area-index for TONG-IL variety was a little retarded than that for PAL-TAL variety, it appeared about 60∼80 days after transplanting. The peaks of the evapotranspiration coefficient and the weight of dry matters at each growth stage were overlapped at about the same time and especially in the later stage of growth, the leaf-area-index, the evapotranspiration coefficient and the weight of dry matters for TONG-IL variety showed a tendency to be larger then those for PAL-TAL variety. 4) The evaporation coefficient at each growth stage for TONG-IL and PAL-TALvarieties was decreased and increased with the increase and decrease in the leaf-area-index, and the evaporation coefficient of TONG-IL variety had a little larger value than that of PAL-TAL variety. 5) Meteorological factors (especially pan evaporation) had a considerable influence to the evapotranspiration, the evaporation and the transpiration. Under the same meteorological conditions, the evapotranspiration (ET) showed a increasing logarithmic function of the weight of dry matters (x), while the evaporation (EV) a decreasing logarithmic function of the weight of dry matters; 800kg/10a x 2000kg/10a, ET=al+bl logl0x (bl>0) EV=a2+b2 log10x (a2>0 b2<0) At the base of the weight of total dry matters, the evapotranspiration and the evaporation for TONG-IL variety were larger as much as 0.3∼2.5% and 7.5∼8.3% respectively than those of PAL-TAL variety, while the transpiration for PAL-TAL variety was larger as much as 1.9∼2.4% than that for TONG-IL variety on the contrary. At the base of the weight of rough rices the evapotranspiration and the transpiration for TONG-IL variety were less as much as 3.5% and 8.l∼16.9% respectively than those for PAL-TAL variety and the evaporation for TONG-IL was much larger by 11.6∼14.8% than that for PAL-TAL variety. 6) The evapotranspiration coefficient, the evaporation coefficient and the transpiration coefficient and the transpiration coefficient were affected by the weight of dry matters much more than by the meteorological conditions. The evapotranspiratioa coefficient (ETC) and the evaporation coefficient (EVC) can be related to the weight of dry matters (x) by the following equations: 800kg/10a x 2000kg/10a, ETC=a3+b3 logl0x (b3>0) EVC=a4+b4 log10x (a4>0, b4>0) At the base of the weights of dry matters, 800kg/10a∼2000kg/10a, the evapotranspiration coefficients for TONG-IL variety were 0.968∼1.474 and those for PAL-TAL variety, 0.939∼1.470, the evaporation coefficients for TONG-IL variety were 0.504∼0.331 and those for PAL-TAL variety, 0.469∼0.308, and the transpiration coefficients for TONG-IL variety were 0.464∼1.143 and those for PAL-TAL variety, 0.470∼1.162. 7) The evapotranspiration ratio, the evaporation ratio (the ratio of the evaporation to the weight of dry matters) and the transpiration ratio were highly affected by the meteorological conditions. And under the same meteorological condition, both the evapotranspiration ratio (ETR) and the evaporation ratio (EVR) showed to be a decreasing logarithmic function of the weight of dry matters (x) as follows: 800kg/10a x 2000kg/10a, ETR=a5+b5 logl0x (a5>0, b5<0) EVR=a6+b6 log10x (a6>0 b6<0) In comparison between TONG-IL and PAL-TAL varieties, at the base of the pan evaporation of 343mm and the weight of dry matters of 800∼2000kg/10a, the evapotranspiration ratios for TONG-IL variety were 413∼247, while those for PAL-TAL variety, 404∼250, the evaporation ratios for TONG-IL variety were 197∼38 while those for PAL-TAL variety, 182∼34, and the transpiration ratios for TONG-IL variety were 216∼209 while those for PAL-TAL variety, 222∼216 (Ref. to table IV-23, table IV-25 and table IV-26) 8) The accumulative values of evapotranspiration intensity and transpiration intensity for both PAL-TAL and TONG-IL varieties were almost constant in every climatic year without the affection of the weight of dry matters. Furthermore the evapotranspiration intensity appeared to have more stable at each growth stage. The peaks of the evapotranspiration intensity and transpiration intensity, for both TONG-IL and PAL-TAL varieties, appeared about 60∼70 days after transplanting, and the peak value of the former was 128.8${\pm}$0.7, for TONG-IL variety while that for PAL-TAL variety, 122.8${\pm}$0.3, and the peak value of the latter was 152.2${\pm}$1.0 for TONG-IL variety while that for PAL-TAL variety, 152.7${\pm}$1.9 (Ref.to table IV-27 and table IV-28) 9) The K-value in Blaney & Criddle formula was changed considerably by the meteorological condition (pan evaporation) and related to be a increasing logarithmic function of the weight of dry matters (x) for both PAL-TAL and TONG-L varieties as follows; 800kg/10a x 2000kg/10a, K=a7+b7 logl0x (b7>0) The K-value for TONG-IL variety was a little larger than that for PAL-TAL variety. 10) The peak values of the evapotranspiration coefficient and k-value at each growth stage for both TONG-IL and PAL-TAL varieties showed up about 60∼70 days after transplanting. The peak values of the former at the base of the weights of total dry matters, 800∼2000kg/10a, were 1.14∼1.82 for TONG-IL variety and 1.12∼1.80, for PAL-TAL variety, and at the base of the weights of rough rices, 400∼1000 kg/10a, were 1.11∼1.79 for TONG-IL variety and 1.17∼1.85 for PAL-TAL variety. The peak values of the latter, at the base of the weights of total dry matters, 800∼2000kg/10a, were 0.83∼1.39 for TONG-IL variety and 0.86∼1.36 for PAL-TAL variety and at the base of the weights of rough rices, 400∼1000kg/10a, 0.85∼1.38 for TONG-IL variety and 0.87∼1.40 for PAL-TAL variety (Ref. to table IV-18 and table IV-32) 11) The reasonable and practicable methods that are applicable for calculating the evapotranspiration of paddy rice in our country are to be followed the following priority a) Using the evapotranspiration coefficients based on an expected yield (Ref. to table IV-13 and table IV-18 or Fig. IV-13). b) Making use of the combination method of seasonal evapotranspiration coefficient and evapotranspiration intensity (Ref. to table IV-13 and table IV-27) c) Adopting the combination method of evapotranspiration ratio and evapotranspiration intensity, under the conditions of paddy field having a higher level of expected yield (Ref. to table IV-23 and table IV-27). d) Applying the k-values calculated by Blaney-Criddle formula. only within the limits of the drought year having the pan evaporation of about 450mm during paddy field period as the design year (Ref. to table IV-32 or Fig. IV-22).