• Korean Journal of Agricultural Science
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• v.36 no.2
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• pp.167-177
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• 2009

This study was conducted to characterize the DGAT1 gene in Korean Holstein dairy cattle population and examine the relationship of DGAT1 polymorphisms with milk yield and milk fat yield for the genetic improvement of Korean Holstein dairy cattle. Results indicated that the 411 bp PCR products were successfully amplified by DGAT1 specific primers. Sequence analysis indicated that the DGTA1 Q allele had AUG (Lysine, K) nucleotide sequences in 216-218 bp and q allele had GCG (Alanine, A) sequences in the same position. Nucleotide sequence homology between the DGAT1 sequences generated in this study showed 100% homology with bovine DGAT1 sequences in the NCBI database. The genotype frequencies of DGAT1 QQ, Qq, and qq were 16.43, 36.43, and 47.14%, respectively, in Korean Holstein dairy cattle population. The observed Q and q allele frequencies were 0.35 and 0.65, respectively. Statistically significant (P<0.05) results were identified for milk yield and milk fat yield for the DGAT1 genotypes. The Qq genotype Holsteins have significantly higher milk yield and milk fat yield than those of the QQ and qq genotype Holsteins(P<0.05).

• Horticultural Science & Technology
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• v.29 no.4
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• pp.366-373
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• 2011

To analyze the genetic relationships among Korean potato cultivars and to develop cultivar identification method using DNA markers, we carried out genotyping using simple sequence repeats (SSR) analysis and developed multiplex-SSR set. Initially, we designed 92 SSR primer combinations reported previously and applied them to twenty four Korean potato cultivars. Among the 92 SSR markers, we selected 14 SSR markers based on polymorphism information contents (PIC) values. PIC values of the selected 14 markers ranged from 0.48 to 0.89 with an average of 0.76. PIC value of PSSR-29 was the lowest with 0.48 and PSSR-191 was the highest with 0.89. UPGMA clustering analysis based on genetic distances using 14 SSR markers classified 21 potato cultivars into 2 clusters. Cluster I and II included 16 and 5 cultivars, respectively. And 3 cultivars were not classified into major cluster group I and II. These 14 SSR markers generated a total of 121 alleles and the average number of alleles per SSR marker was 10.8 with a range from 3 to 34. Among the selected markers, we combined three SSR markers, PSSR-17, PSSR-24 and PSSR-24, as a multiplex-SSR set. This multiplex-SSR set used in the study can distinguish all the cultivars with one time PCR and PAGE (Polyacrylamide gel electrophoresis) analysis and PIC value of multiplex-SSR set was 0.95.

• Journal of Animal Science and Technology
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• v.44 no.2
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• pp.181-190
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• 2002

This study was performed to determine sequences of the mt DNA D-loop region, including $tRNA^{Pro}$ and $tRNA^{Pre}$ and to analysis sequence variation polymorphism in Korean cattle. The resulting sequencies were compared with previously published sequences for other cattle breeds(GenBank J01394). The PCR was used to amplify an 1142bp between nucleotides 15061 and 404 within the D-loop region of mt DNA using specific primers. Korean cattle showed 24 polymorphic sites by nucleotide substitutions and insertions of single base pairs. About 50% of polymorphic sites were found in positions 16042 to 16122 with the most variable region. Among these polymorphic sites, variations at 16055, 16230 and 16260 bp were detected as new sequence variants in Korean cattle. These specific polymorphic sites have not been reported in the Japanese black cattle and European cattle. Therefore, mt DNA variants in the D-loop region may be used as genetic markers for specifying Korean cattle. The frequencies of positions 169, 16302, 16093, 16042, 16119 with a high level of sequence polymorphism were 0.81, 0.56, 0.56, 0.50 and 0.43, respectively. In comparison of genetic distances, Korean cattle showed the more closely to European cattle as Bos taurus than Bos indicus such as African and India breeds. In conclusion, these mt DNA sequence polymorphisms in the D-loop region for Korean cattle may be useful for the analysis of cytoplasmic genetic variation and associations with economic important traits and genetic analysis of maternal lineage.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.3
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• pp.169-175
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• 2016

Saccharin (o-benzoic sulfimide) is the first artificial and non-caloric sweetener that was first synthesized in 1879. In this study, we examined the biological activity of saccharin against various human cancer cell lines and human bone marrow-derived mesenchymal stem cells. A viability assay based on the conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was performed to test for the cytotoxicity of saccharin about the four human cancer cell lines (H460, H157, A549 and SKOV3), one murine cancer cellline (Raw264.7), and MSCs. In order to find the differentially expressed gene in saccharin-treated MSCs against untreated MSCs, we performed annealing control primer (ACP)-based differential display reverse transcriptionp-olymerase chain reaction (DDRT-PCR). All tested cells were treated with saccharin at various concentrations (0.0, 4.8, 7.2, 9.6, 12.0, 14.4 mg/mL) for 48 hr. The number of metabolically active cancer cells decreased when treated with the saccharin at various concentrations for 48 hr as compared with the untreated cells. The decrease in cell survival was more evident with increasing concentrations of saccharin. Moreover, novel candidate genes, which were differentially expressed in MSCs in response to saccharin, were identified in 16 bands on 2% agarose gel. This revealed 16-7 up-regulated and 9 down-regulated-differentially expressed genes indicated by arrows. One of these candidate genes was a FK506-binding protein gene. The functional roles of FK506 binding proteins, with respect to the activities of stem cell proliferation, were not characterized. Further studies are required to get a better understanding of FK506-binding proteins in its roles in increasing stem cell proliferative activities from using saccharin.

• Journal of Animal Science and Technology
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• v.50 no.5
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• pp.641-648
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• 2008

There has been great success for making transgenic animals using somatic cell nuclear transfer(SCNT) up to this time. However, the success rates of the production of live transgenic animals are still very low. The current research has been carried out for delineation of differentially expressed genes between SCNT and normal placenta in cattle. In the present observations, high expression has been observed for CTSZ, LOC509426 and ELF1 genes in normal placenta. On the other hand, TIMP2, PAG1B, PAG-21, LOC782894, SERPINB6 and mKIAA2025 protein were highly expressed in SCNT placenta. Five genes, which were highly expressed in SCNT placenta, have been further investigated using semi-quantitative real-time PCR. The results were similar to that we observed using ACP. In the future, all genes affecting the SCNT and normal placenta have to be discovered and their networks will be fully investigated. The genes were identified in this study would be great help for identifying differential gene expressions in SCNT placenta.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.2
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• pp.105-110
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• 2004

Objectives: Despite severe oligospermia, males with Y chromosome microdeletion can achieve conception through ICSI (Intracytoplasmic Sperm Injection). However, ICSI may not only result in the transmission of microdeletions but also the expansion of deletion to the offspring. The purpose of this study was to screen vertical transmission, expansion of microdeletions and de novo deletion in male fetuses conceived by ICSI. Materials and Methods: A total of 32 ICSI treated patients with their 33 (a case of twin) male fetuses conceived by ICSI were used to make this study group. Sequence-tagged sites (STSs)-based PCR analyses were performed on genomic DNA isolated from peripheral blood of fathers and from the amniocytes of male fetuses. Ten primer pairs namely, sY134, sY138, MK5, sY152, sY147, sY254, sY255, SPGY1, sY269 and sY158 were used. The samples with deletions were verified at least three times. Results: We detected a frequency of 12.5% (4 of the 32 patients) of microdeletions in ICSI patients. In 4 patients with detected deletions, two patients have proven deletions on single STS marker and their male fetuses have the identical deletion in this region. Another two patients have two and three deletions, but their male fetuses have more than 3 deletions which include deletions to their father's. Meanwhile, seven male fetuses, whose fathers were analyzed to have all 10 STS markers present, have deletions present in at least one or more of the markers. Conclusions: Although the majority of deletions on the Y chromosome are believed to arise de novo, in some cases a deletion has been transmitted from the fertile father to the infertile patient. In other cases the deletion was transmitted through ICSI treatment, it is likely that one sperm cell is injected through the oocyte's cytoplasm and fertilization can be obtained from spermatozoa. Our tests for deletion were determined by PCR and our results show that the ICSI treatment may lead to vertical transmission, expansion and de novo Y chromosome microdeletions in male fetuses. Because the sample group was relatively small, one should be cautious in analyzing these data. However, it is important to counsel infertile couples contemplating ICSI if the male carries Y chromosomal microdeletions.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.6
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• pp.875-880
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• 2012

This study was conducted to investigate the level of contamination of pathogenic Escherichia (E.) coli in 50 types of red pepper powders collected domestically. Pathogenic E. coli was confirmed using real-time PCR to confirm the 4 types of EAEC, EPEC, EHEC and ETEC. One sample out of 50 was contaminated with pathogenic E. coli. The type of pathogenic E. coli detected in the sample was EAEC. This study was also conducted to determine the effect of alcohol treatment on the reduction of E. coli populations in red pepper powder. The amount of E. coli in the control was $1.2{\times}10^6$ cfu/mL. The amount of E. coli in 10 minutes immersion treatment with 10% alcohol was $1.1{\times}10^6$ cfu/mL. In samples treated with over 20% alcohol, E. coli was not detected. This showed that 10 minutes of immersion in over 20% alcohol might be effective to reduce E. coli. This study was also conducted to determine the effect of UV irradiation on E. coli reduction. The number of E. coli in the control group was $5.0{\times}10^5$ cfu/mL. However, the number of E. coli in 45 min of the UV irradiated sample decreased to $1.0{\times}10^3$ cfu/mL, by $10^2$ cfu/mL. In contrast, E. coli was not detected in an over 60 min UV irradiated sample in $10^{-2}dilution$. This study showed that over 20% alcohol treatment and UV irradiation for 60 min was effective to control E. coli in red pepper powder.

• Clinical and Experimental Pediatrics
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• v.48 no.2
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• pp.197-203
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• 2005

Purpose : Changes in metalloproteinases(MMP) activity have been demonstrated in several disease states, including rheumatoid arthritis and tumor metastasis. More importantly, increased myocardial MMP activity has been reported to occur in both clinical and experimental forms of dilated cardiomyopathy. There was no report about MMP in adriamycin(ADR)-induced cardiomyopathy. The purpose of this study was to investigate gene expression of MMP and tissue inhibitor of metalloproteinases(TIMP) in ADR-induced cardiomyopathy and clarify the relationship between MMP and cytokines. Methods : Male Sprague-Dawley rats were divided into two groups. The first group was control. The second group was given intraperitoneal injections of ADR(5 mg/kg) twice a week over two weeks. Serum concentrations of MMP, TIMP, interleukin(IL)-6 and tumor necrosis factor(TNF)-${\alpha}$ were measured. RNA extraction was performed from frozen rat hearts. Reverse transcription polymerase chain reaction(RT-PCR) was employed. cDNA Microarray analysis was performed by using a set of 5,184 sequence-verified rat cDNA clones. Results : Serum MMP and TIMP levels were not significantly different between the two groups. IL-6 was $36.8{\pm}2.8pg/mL$ and TNF-${\alpha}$ $2.2{\pm}2.7pg/mL$ in the ADR group. They were significantly higher than in the control group. Serum MMP correlated significantly with TNF-${\alpha}$(r=0.41, P<0.05). There was no gene expression of MMP, IL-6 or TNF-${\alpha}$ in the hearts of both groups. Gene expression of TIMP was significantly depressed in the hearts of the ADR group. Conclusion : These results suggested a potential role for TNF-${\alpha}$ in the regulation of extracellular matrix remodeling in ADR induced cardiomyopathy. Rapid screening of multiple decreased gene expression by DNA chip may be a useful diagnostic test to detect early cardiac injury before developing ADR induced cardiomyopathy.

• Journal of Korean Society of Forest Science
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• v.90 no.1
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• pp.155-160
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• 2001

Changes in genetic diversity after seed-tree practice were simulated in test plantation of tulip tree at Sokdal-ri, Banwol-myon, Hwasung-gun, Kyungki Provence. I-SSR markers were analyzed in a total of 305 tulip trees. A total of 89 amplicon variants were observed by PCR with 9 I-SSR primers. Genetic diversity for 305 trees was relatively high (S. I. =0.4532). Individuals originated from the seed orchards in U.S.A. showed the highest level of genetic diversity (0.4530), those from Anyang showed the medium level (0.4152), and those from Cheonbuk showed the lowest (0.3929). Simulation of seed-tree practice accompanied by 2 consecutive thinnings was performed on the basis of morphological characteristics and planted distances of the individual trees, which left 37 trees as candidates for seed-trees. Decreasing rate of genetic diversity within seed sources was greatest for individuals from Cheonbuk (28.3%), moderate for those from Anyang (16.3%), and smallest for those from U.S.A. (8.0%). In spite of little difference in decreasing rate of individuals for the 3 seed sources (87.5~88.2%), large difference in decreasing rate of genetic diversity within seed sources might be due to difference in number of mother trees for the 3 seed sources. For example, whereas individuals originated from the seed orchard in U.S.A., which showed the smallest decreasing rate of genetic diversity, might be originated from relatively large number of mother trees, those from Anyang and Cheonbuk might be originated from relatively small number of mother trees. Although mean of 17.5% of the genetic diversity within seed sources was decreased through 2 consecutive thinnings, a decrease in genetic diversity for the whole individuals (37 trees) was only 6.1%. This observation suggests that the seed-tree practice on the basis of the criteria established in the present study may not result in great reduction in overall genetic diversity of the progenies.

• Journal of the Korean Society of International Agriculture
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• v.30 no.4
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• pp.347-356
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• 2018

Genetically modified (GM) crops have been increased continuously over the world and concerns about the potential risks of GM crops have also been increasing. Even though GM crops have not been cultivated commercially in Korea, it should be necessary to develop the safety assesment technology for GM crops. In this study, we investigated the influence of heading date difference on gene flow from GM to non-GM rice. In the experimental plot design, The PAC GM rice was placed in the center as a pollen donor and non-GM rice were placed in eight directions as pollen receivers. Five pollen receiver rice cultivars were Unkawng, Daebo, Saegyejinmi, Nakdong-byeo, and Ilmi which had different flowering times. A total of 266,436, 300,237, 305,223, 273,373, and 290,759 seeds were collected from Unkawng, Daebo, Saegyejinmi, Nakdong, and Ilmi, respectively, which were planted around PAC GM rice. The GM${\times}$non-GM hybrids were detected by repeated spraying of herbicide and PAT immunostrip assay. Finally, the hybrids were confirmed by PCR analysis using PAC gene specific primer. The hybrids were found in Nakdong-byeo which had the same heading date with PAC GM rice. The hybridization rate was 0.0007% at Nakdong-byeo plot. All of GM${\times}$non-GM hybrids were located within 2 m distance from the PAC GM rice zone. The physiological elements including rice heading date were found to be important factors to determine GM?rice out crossing rate with GM rice. Consideration should be taken into for many factors like the physiological elements of field heading date of rice cultivars to set up the safety management guideline for prevention of GM rice gene flow.