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Development of Bovine Embryos after Vitrified-Thawed with Electron Microscope Grid and Open Pulled Straws

  • Lee, Y. J.;D.H. Ko;Lee, H. T.;K. S. Chung
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.76-76
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    • 2001
  • The objective of this study was to optimize the vitrification method of in vitro produced bovine embryos. Thus, in vitro produced embryos at 8 cell, morula and blastocyst stages were vitrified on electron microscope grids (EM grids) or in open pulled straws (OPS) with EG5.5 (5.5 M ethylene glycol, 1.0 M sucrose and 10% FBS in m-DPBS medium) freezing solution and their survival rates after thawing were compared. The embryos on EM grids or in OPS were briefly exposed to EG5.5 freezing solution and plunged directly into liquid nitrogen within 30 to 35 sec. Post-thawed embryos were serially diluted in 0.5, 0.25 and 0.125 M sucrose in m-DPBS, each for 1 min, and then cultured in CRI aa medium supplemented with 10% FBS. Embryonic survival rate was assessed as re-expanded and hatched rates of those embryos after warming. The rates of re-expansion embryos did not significantly different between EM grid (8 cell: 42.10%, morula: 66.66% and blastocyst: 77.08%) and OPS (8 cell: 47.36%, morula: 61.90% and blastocyst: 83.33%) methods. In addition, the hatched rates in EM grid (8 cell: 31.57%, morula: 57.14% and blastocyst: 72.91%) were similar to those in OPS (8 cell. 34.21%, morula: 50.00% and Blastocyst: 77.08%). Interestingly, even at the same blastocyst stage, the in vitro survival of day 7 embryos (EM grid: 79.48 and OPS: 87.18%) was higher than those of day 8 embryos (EM grid: 72.10 and OPS: 82.06%). The total cell number of blastocyst developed in vitro after vitrification was examined with Hoechst 33342 staining to compare the embryo quality among different treatment groups. The total cell number of blastocyst was not significantly different between vitrified groups (EM grid: 162.4$\pm$8.0 and OPS: 158.4$\pm$7.1) and unvitrified control (168.0$\pm$5.6). These results indicate that both vitrification containers can provide the high rate of embryo survival. Moreover, the OPS container may not need a cap to protect the container from floating after immersion in L$N_2$. Therefore, this study suggest that bovine embryos can be cryopreserved easily, effectively and successfully by vitrification method using EM grid or OPS with EG5.5 freezing solution. In the future, the Pregnancy rate would be investigated after transfer of our vitrified embryos into the appropriated recipients.

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Transitional Patterns of Vegetation in Reclaimed Land Applied with Solidified Sewage Sludge (하수슬러지 고화물을 처리한 매립예정 간척지토양의 잡초발생 양상변화)

  • Um, Kyoung Ran;Jang, Yun-Hui;An, Gi Hong;Cha, Young-Lok;Yu, Gyeong-Dan;Lee, Ji-Eun;Moon, Youn-Ho;Ahn, Joung Woong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.3
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    • pp.381-387
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    • 2015
  • This study was firstly conducted to investigate changes of vegetation and soil characteristics in reclaimed land applied with solidified sewage sludge for the cultivation of bioenergy crops. Each vegetation survey site was approximately $15m{\times}3m$ on the inside of each experimental plot that consisted of 50% (A-1), 30% (A-2), 15% (A-3), and 5% (A-4) mixture of solidified sewage sludge, and original reclaimed soil (ORS). After the application of solidified sewage sludge, we monitored the changes of vegetation and soil properties for three years. In first year, soil pH, electrical conductivity (EC) and exchangeable $Ca^{2+}$ content was 9.4~10.8, $9.10{\sim}14.41\;dS\;m^{-1}$, and $62.1{\sim}204.2\;cmol\;kg^{-1}$, respectively, while three years later, it decreased to 8.1~8.4, $1.65{\sim}5.98\;dS\;m^{-1}$, and $21.9{\sim}43.1\;cmol\;kg^{-1}$, respectively. These results indicated that several of soil chemical elements which have nagative impacts on the plant growth in the plots of mixtures of solidified sewage sludge, steadily declined as the years go by. The vegetations in each survey site were recorded as 6 families and 12 species in 2014, while the vegetations were not occurred at all survey sites in 2012, and only halophytes as Phragmites australis and Suaeda asparagoides were observed in 2013. Diversity of vegetation, which was calculated by shannon index (H'), increased as the season progressed at each experimental plot applied with solidified sewage sludge. In original reclaimed soil, however, there was showed the high community similarity of vegetation due to the fact that P. australis and S. asparagoides were only occurred for survey periods.

Acid Tolerance of the Acid-Resistant Mutant of Leuconostoc paramesenteroides Improved for Kimchi Starter. (김치 Starter용으로 개량된 Leuconostoc paramesenteroides의 내산성 변이주가 갖는 내산성 특성)

  • 김영찬;정은영;김은해;정대현;최태부;권태종;강상모
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.275-282
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    • 1998
  • To investigate the increased acid tolerance of a acid-resistant mutant Leuconostoc paramesenteroides P-100 improved as a kimchi starter, proton permeability, ATPase acitivity, glycolysis activity, $Mg^2$sup +/ releasement, and membrane fatty acid composition were studied and comprised with its wild type Leuconostoc paramesenteroides Pw. In the proton permeability experiment, the maximum values of the average half time (t$\_$1/2/) of pH equilibration through the cell membrane of the Pw and the P-100 were about 6.4 min and 7.8 min in 150 mM KCI solution, respectively. In the 3% NaCl solution, the t$\_$1/2/ values of the Pw and the P-100 were 5.5 min and 6.9 min, respectively. The values and pHs of maximal specific activities of ATPase originated from the Pw and the P-100 were 0.5 unit/mg protein and 0.78 unit/mg protein at pH 6.0, respectively. The result of pH dependence of glycolysis showed that the P-100 had higher activities than that of Pw except at pH 7.0. The releases of magnesium from the Pw and the P-100 were observed about 54.5% and 23.2% at pH 4.0 after 2 hours, respectively. The results of comparison of membrane fatty acid composition of the Pw with the P-100 showed that C$\_$8:0/, C$\_$9:0/, C$\_$10:0/, C$\_$11:0/, C$\_$18:0/, and C$\_$19:0,cyclo/ were major different fatty acids between two strains and the content of C$\_$18:1/, and C$\_$19:0,cyclo/ were 2.8%, N.D (not detected) in the Pw and 0.4%, 2.3% in the P-100. These results indicated that acid tolerance of the P-100 was significantly improved in comparison with its wild type Pw.

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INVESTIGATION OF THE EFFECT OF AN ANTIBIOTIC "P" ON POTATOES ("감자에 대한 항생제(抗生劑) 피마리신의 통계적(統計的) 효과(效果) 분석(分析)")

  • Kim, Jong-Hoon
    • Journal of Korean Society for Quality Management
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    • v.5 no.2
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    • pp.59-120
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    • 1977
  • An antibiotic 'P', which is one of the products of the Gist Brocades N. V. is being tested by its research department as fungicide on seed-potatoes. For this testing they designed experiments, with two control groups, one competitor's product, eight formulations of the antibiotic to be tested in different concentrations and one mercury treatment which can not be used in practice. The treated potatoes were planted in three different regions, where bifferent conditions prevail. After several months the harvested potatoes are divided in groups according to their diameter, potato illness is analysed and counted. These data were summarised in percentage and given to us for Analysis. We approached and analysed the data by following methods: a. Computation of the mean and standard deviation of the percenage of good results in each size group and treatment. b. Computation of the experimental errors by substraction of each treatment mean from observed data. c. Description of the frequency table, plotting of a histogram and a normal curve on same graph to check normality. d. Test of normality paper and chi-sqeare test to check the goodness of fit to a normal curve. e. Test for homogeneity of variance in each treatment with the Cochran's test and Hartley's test. f. Analysis of Variance for testing the means by one way classifications. g. Drawing of graphs with upper and lower confidence limits to show the effect of different treatments. h. T-test and F-test to two Control mean and variance for making one control of Dunnett's test. i. Dunnett's Test and calculations for numerical comarision of different treatments wth one control. In region R, where the potatoes were planted, it was this year very dry and rather bad conditions to grow potatoes prevailed during the experimental period. The results of this investigation show us that treatment No.2, 3 and 4 are significantly different from other treatments and control groups (none treated, just like natural state). Treatment no.2 is the useless mercury formulation. So only No. 3 and 4, which have high concentrations of antibiotic 'P', gave a good effect to the potatoes. As well as the competitors product, middle and low concentrated formulations are not significantly different from control gro-ups of every size. In region w, where the potatoes got the same treatments as in region R, prevailed better weather conditions and was enough water obtainable from the lake. The results in this region showed that treatment No. 2, 3, 4, and 5 are Significantly different from other treatments and the control groups. Again No.2 is the mercury treatmentin this investigation. Not only high concentrated formulation of antibiotic 'P', but also the competitor's poroduct gave good results. But, the effect of 'P', was better than the competitors porduct. In region G, where the potatoes got the same treatments as in the regions R and w. and the climate conditions were equal to region R, the results showed that most of the treatments are not significantly different from the control groups. Only treatment no. 3 was a little bit different from the others. but not Significantly different. It seems to us that the difference between the results in the three regions was caused by certain conditions like, the nature of the soil the degres of moisture and hours of sunshine, but we are not sure of that. As a conclusion, we can say that antibiotic 'P' has a good effect on potatoes, but in most investigations a rather high concentration of 'P' was required in formulations.

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Establishment of Bovine Ovum Bank : I. Full Term Development of Vitrified In Vitro Matured Hanwoo (Korean Cattle) Oocytes by Minimum Volume Cooling (UC) Method (소 난자 은행 설립 : I. MVC 방법으로 초자화 동결된 한우 미성숙 난자의 개체 발생능 조사)

  • Kim, E.Y.;Kim, D.I.;Rhee, M.G.;Weon, Y.S.;Nam, H.K.;Lee, K.S.;Park, S.Y.;Park, E.M.;Yoon, J.Y.;Heo, Y.T.;Cho, H.J.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Korean Journal of Animal Reproduction
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    • v.25 no.1
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    • pp.1-7
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    • 2001
  • This study was to test whether in vitro matured Hanwoo oocytes can be successfully cryopreserved by a new vitrification procedure using MVC method. For the vitrification, oocytes were pretreated in 10% ethylene glycol (EG10) for 5~10 min, exposed in EG30 for 30 sec, each oocyte was individually put on the inner wall of 0.25 $m\ell$ straw, and then straws were directly plunged into L$N_2$. Thawing was taken by 4-step procedures 〔1.0 M sucrose (MS), 0.5 MS, 0.25 MS, and 0.125 MS〕 at 37$^{\circ}C$. In vitro developmental capacity (survival, cleavage ($\geq$2-cell) and blastocyst rates) in vitrified group was no significant difference compared to that in other treatment groups (exposed; 100.0, 74.4, 32.3% and control; 100.0, 78.3, 36.3%): high mean percentage of oocytes (91.2%) was survived, 69.4% of them were cleaved and 27.9% of cleaved embryos were developed to blastocyst. Especially, after transfer of in vitro developed embryos in vitrified group, four of six recipient animals were pregnant and three of them were ongoing-pregnant by manual palpation at 250 days after transfer. This result demonstrates that MVC method is very appropriate freezing method for the Hanwoo in vitro matured oocytes and that ovum bank can be maintained efficiently by MVC cryopreservation method.

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Evaluation of Sodium Intake and Relationship between Sodium Intake and the Bone Mineral Density of Female University Students (중부 지역 여대생에서 음식섭취빈도조사지를 이용한 나트륨 섭취량 평가 및 나트륨 섭취와 골밀도와의 관련성 조사)

  • Bae, Yun-Jung;Yeon, Jee-Young
    • Journal of the East Asian Society of Dietary Life
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    • v.21 no.5
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    • pp.625-636
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    • 2011
  • The purpose of this study was to evaluate the relationship between bone health and sodium intake in female university students using a dish frequency questionnaire (DFQ 125), anthropometric checkups, food records for 3 days, and ultrasound measurement of calcaneus bone mineral density. Subjects were divided into two groups: normal (n=196) and osteopenia (n=52). There were no significant differences in age or height between the two groups. The average weight, body mass index, and body fat in the normal group were significantly higher than in the osteopenia group. The sodium intake of DFQ was positively correlated with the sodium intake of 3 days of dietary records (p=0.0003). There were no significant differences in the sodium intake between the two groups from DFQ. The dishes were ranked by sodium intake: kimchies were 17.68%, noodles and mandu were 16.36%, stews were 13.69%, main dishes such as meat, egg, and beans were 11.47%, and fish and shellfish were 11.07%. The frequency of eating noodles and mandu (p=0.0116), stews (p=0.0008), kimchies (p=0.0482), fish and shellfish (p=0.0362), vegetables (p=0.0064) and seasoning (p=0.0347) were negatively associated with bone mineral density. Bone health was not significantly different with increasing quartiles of sodium intake. As excessive sodium intakes may indirectly affect bone mineral density, these results suggest that to prevent osteoporosis, university students needed to be more educated about diets containing less sodium through nutrition education programs.

Nutritional Composition and Physico-chemical Meat Quality Properties of Korean Hanwoo Bull Beef (한우수소의 부위별 영양성분조성 및 이화학적 육질특성)

  • Cho, S.H.;Park, B.Y.;Kim, J.H.;Choi, Y.H.;Seong, P.N.;Chung, W.T.;Chung, M.O.;Kim, D.H.;Ahn, C.N.
    • Journal of Animal Science and Technology
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    • v.49 no.6
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    • pp.871-880
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    • 2007
  • This study was conducted to investigate the relationship between the socio-demographic factors and the Korean consumers’ palatability evaluation and to collect the sensory information for development of prediction palatability model of Hanwoo beef. Ten cuts〔Abjin (short plate), Bosup (top sirloin), Cheggt (striploin), Dngsim (loin), Guri (chuck tender), Hongduke (eye of round), Moksim (chuck roll), Sulgit (bottom round), Udoon (top round), Yangi (brisket)〕 were separated from 10 Hanwoo bulls beef(~24 months old) and prepared with the same manner for 3 different cooking methods such as boiling, grilling and roasting. The cooked beef samples were served to 650 consumers recruited from Seoul, Joongbu, Honam and Youngnam locations and evaluated tenderness, juiciness, flavor and overall acceptability. The living location, age, gender, occupation, eating habit and cut were significantly related with the sensory properties when bull beef were cooked as boiled cooking(p<0.05). In grill cooking, only occupation, income and cut had the significant effect on the overall acceptability of Hanwoo bull beef(p<0.01). When bull beef were cooked as Korean traditional roast cooking, consumer’s sensory properties were significantly different by the living location, age, occupation, income, eating habit of consumers and cut(p<0.05). Also, results from the principal component analysis showed that palatability scores of Korean consumers were decided with different contribution scores of tenderness, juiciness, flavor and overall acceptability of cut depending on cooking methods. In conclusion, Korean consumers’ palatability for Hanwoo bull beef were related with the socio-demographic factors and the sensory results were different by cut and cooking methods.

Effect of reaction temperature on the particle size and crystal shape of precipitated calcium carbonate (반응온도가 침강성탄산칼슘의 입도 및 형상에 미치는 영향)

  • 송영준;박찬훈;조동성
    • Resources Recycling
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    • v.4 no.1
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    • pp.38-45
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    • 1995
  • The objective of this study was to investigate the effect of temperature on the formation of CaCO, polymorphs(i.e.,calcite, aragonite, vaterite) and on the crystal shape of CaCO,.The reaction systems were rnvestigated at the temperature range of 2.0%-85.3r, at the fixed cmditions ofconcentration and pressure, 2X10-' M, atomospheric pressure, respectively.The reaction systems studied include a Ca(HCO.,),-Air bubble, O Ca(OH)s-CO,, @ Ca(OH),-H,CO, ,Ca(OH1,-Na>CO,, O Ca(OH),-K,CO,, @ Ca(OH),-(NH,),CO,, D CaC1,-Na,CO,, CaC1,-K3C03, 8 CaC1,-(NH,,),CO,, 0 Ca(N0,X-Na,CO,, 03 Ca(N0,X-QCO,. 0 Ca(NO,),-(NH,XCO,. The results obtained are summarizedas follows:Calcite is formed at the temperature range of 2t-80"C and the highest calcite yield was obtained at 30%.Aragonite begins to be formed at the temperature range of 41.0%-53.0%. and the higher temperature is thehigher yield is obtained. pH of the reaction system affect the yield of aragonite, and the yield reaches the highestpercentage at the pH range of 10.0-11.0, and at the conditions of pH 12.3 or over, aragonite is scarcely formed.Vaterlle is fnrmed at the temperature range of 40.0% or less, and transites utterly to calcite within 10-60mmutes in the case of bemg residenced in mother liqmd which C1 is not contained, and within 140hours inthe case of containing CI-.s in the case of containing CI-.

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Effects of Different Microbial Culture Supplements on In vitro and In situ Ruminal Fermentation Characteristics of Italian ryegrass Silage (미생물제 처리에 의한 이탈리안 라이그라스 사일리지의 In vitro 및 In situ 반추위 발효특성에 미치는 영향)

  • Lim, D.H.;Ki, K.S.;Choi, S.H.;Kim, T.I.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.36 no.4
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    • pp.309-317
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    • 2016
  • The study was conducted to evaluate the effects of microbial culture supplements on ruminal fermentation and fermentative quality of Italian ryegrass silage (IRGS) both in vitro and in situ. Three species of microbes (Lactobacillus casei (LC), Bacillus subtilis (BS), and Saccharomyces cerevisiae (SC)) were used in this study. They were applied to IRGS at 30 days after silage manufacture. Various items were measured using in vitro and in situ incubation technique after each microbial supplement was inoculated into IRGS at $0.5{\times}10^4CFU/g$. In the first experiment, in vitro ruminal fermentation characteristics of IRGS were evaluated at 0, 12, 24, 48, and 72 hours after microbes were inoculated into IRGS. In the second experiment, in situ fermentation characteristics were investigated at 0, 1, 3, and 5 days after the inoculation of each microbial supplement. In vitro ruminal $NH_3-N$ content was significantly (p<0.05) increased in LC-, BS-, and SC-IRGS at 12 hrs post incubation compared to that in control IRGS. In vitro ruminal total VFA concentration and dry matter digestibility (DMD) of IRGS were not significantly difference among LC-, BS-, and SC-IRGS, although they were numerically increased in LC-IRGS than those of the other IRGS. In addition, this study evaluated the fermentation characteristics and in situ DMD of IRGS with the lapse of incubation time up to 5 days. Throughout the incubation times from 1 day to 5 days, the pH value was significantly (p<0.05) lower in BS-, LC-, and SC-IRGS than that in control IRGS. Lactate was significantly (p<0.05) higher, and significantly (p<0.05) butyrate was lower in LC-IRGS than that in other treatments at 0 day. It was higher (p<0.05) in control IRGS than that of BS-, LC-, and SC-IRGS at 1-5 days. In situ DMD tended to increase in BS-, LC-, and SC-IRGS compared to that in control IRGS. Especially, DMD was higher in SC-IRGS than that in other treatments at 0 day. It tended to be higher in LC-IRGS at all incubation time. Taken together, these results suggest that it might be useful to select a microorganism by considering the feeding time of IRGS to ruminants because organic acids and DMD of IRGS were affected by the incubation time of each microorganism with IRG silage, especially for L. casei decreased the content of acetate and butyrate in IRGS.

Investigation of Coat Color Candidate Genes in Korean Cattle(Hanwoo) (한우에서 모색관련 유전자 변이에 관한 연구)

  • Do, K.T.;Shin, H.Y.;Lee, J.H.;Kim, N.S.;Park, E.W.;Yoon, D.H.;Kim, K.S.
    • Journal of Animal Science and Technology
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    • v.49 no.6
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    • pp.711-718
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    • 2007
  • Most cattle breeds have a coat color pattern that is characteristic for the breed. Korean cattle(Hanwoo) has a coat color ranging from yellowish brown to dark brown including a red coat color. Variation in the Hanwoo coat color is likely to be the effects of modified genes segregating within the Hanwoo breed. MC1R encoded by the Extension(E) locus was almost fixed with recessive red e allele in the Hanwoo, but other gene(s) might be affecting the variation of the Hanwoo coat color into yellowish to red brown. We have analyzed a segregation of coat color in the F2 families generated from two Hanwoo bulls(yellowish brown) mated to six F1 dams(black) derived from Hanwoo and Holstein crosses. Segregation of coat color in the offspring found a ratio of 1(yellowish brown) : 1(black) and this ratio indicates that a single gene may play a major role for the Hanwoo coat color. We further investigated SNPs in MC1R, ASIP and TYRP1 loci to determine genetic cause of the Hanwoo coat color. Several polymorphisms within ASIP intron 2 and TYRP1 exons were found but not conserved within the Hanwoo population. However, the segregation of the MC1R e allele was completely associated with the Hanwoo coat color. Based on this information, it is clear that the MC1R e allele is mainly responsible for the yellowish red Hanwoo coat color. Further study is warrant to identify possible genetic interaction between MC1R e allele and other coat color related gene(s) for the variation of Hanwoo coat color from yellowish brown to dark brown. (Key words : Hanwoo, Coat color, SNP, MC1R, ASIP, TYRP1)