• Title/Summary/Keyword: artificial gene

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Discontinuous Percoll Gradients Enrich X-Bearing Porcine Sperms and Female Embryos (불연속 Percoll 원심분리에 의한 돼지 X-정자와 자성배아에 관한 연구)

  • Shim, Dae-Yong;Yoo, Seong-Jin;Kang, Han-Seung;Yoo, Jeong-Min;Lee, Chae-Kwan;Kang, Sung-Goo
    • Development and Reproduction
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    • v.5 no.1
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    • pp.47-52
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    • 2001
  • Predetermination of sex in livestock of offpring is in great demand and is of critical importance to providing for the most efficient production of the animal ariculture. Such a sexing techlology would also enhance the economy of conventional artificial insemination(AI) and aid the porcine industry. The purpose of this study was to evaluate the efficiency of enriching X-bearing porcine sperm using discontinuous percoll gradients and PCR mefhod. Semen was collected from mature boars of proven fertility center (AI center KimHae). Sperm was leaded on the isotonic discontinuous percoll gradient and then it was centrifuged at 120 ${\times}$ g for 20 minutes. After centrifugation, sperm included in each fraction were recovered (7${\times}$10$^6$ sperms/ml) and then sperm genomic DNA was extractedfor the PCR. SRY gene was used to evaluate the ratio between X and Y sperm in the separated fractions. Ju viro ffrtilization wascarried out by adding the unseparated sperm (control) or separated (experimental poop) to the matured oocytes in TCM-199. Embryos for sex determination were obtained at 2 cell stage and then was used for SRY gene amplification. After centrifugation of discontinuous percoll gradient, the most motile sperm was obtained at 95% fiaction (94.4% ${\pm}$ 5.1%, p < 0.01). The PCR analysis evaluated that 30%, 50% and 65% fractions were Y sperm rich, whereas 80% and 95% fractions were X sperm rich. PCR analysis with each porcineembryo showed that 33.3% of control and 66.7% of experimental group were determined as female embryos. In conclusion, in vitro matured oocytes inseminated with sperms (95% fraction) prepared by percoll gradient centrifugation showed high fertilization rates and female embryos than control sperms.

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Fermentative characteristics of yogurt using lactic acid bacteria isolated from Korean traditional fermented food (전통 발효 식품에서 분리한 유산균을 이용한 yogurt 발효특성)

  • Park, Na-Young;Lee, Shin-Ho
    • Food Science and Preservation
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    • v.24 no.5
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    • pp.707-713
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    • 2017
  • The objective of this study was to select yogurt starter from Korean traditional fermented foods. The 2 strains (KM24, KM32) among 50 strains of isolated lactic acid bacteria selected as starter based on milk clotting ability, antimicrobial activity against various pathogens, tolerance in artificial gastric and bile juice and growth in 10 % skimmed milk. The strains were identified as Lacobacillus plantarum (KM32) and Pediococcus pentosacesus (KM24) by 16S rRNA gene sequencing. Viable cell number of yogurt fermented with mixed strains (KM24 and KM32) was 9.66 log CFU/mL after fermentation for 48 h and maintained $10^9CFU/mL$ during fermentation for 72 h at $37^{\circ}C$. The pH and titratable acidity of mixed cultured yogurt were 4.25% and 0.83% after fermentation for 48 h at $37^{\circ}C$, respectively. The physico-chemical characteristics of mixed cultured yogurt after fermentation for 48 h were $38.45{\mu}g/mL$ (polyphenol content), 48.57% (DPPH radical scavenging activity) and 465.40 cp (viscosity), respectively. The mixed cultured yogurt maintained $10^9CFU/mL$ of lactic acid bacteria during storage 10 days at $4^{\circ}C$. The viable cell number of yogurt prepared with mixed culture(KM32+KM24) maintained higher and than that of control (L. casei) during storage. These results indicated the potential use of selected strains (KM32+KM24) isolated from kimchi as a yogurt starter with strong acid tolerance and probiotics properties.

Verification of ET and AI Derived Offspring Using on the Genetic Polymorphisms of Microsatellite and Coat Color Related Genes in Jeju Black Cattle (제주흑우 집단에서 모색 관련 유전자와 microsatellite marker의 다형현상을 이용한 수정란이식 및 인공수정 유래 후대우 검증)

  • Han, Sang-Hyun;Ko, Jin-Cheul;Kim, Young-Hoon;Kim, Nam-Young;Kim, Jae-Hwan;Ko, Moon-Suck;Jeong, Ha-Yeon;Cho, In-Cheol;Yang, Young-Hoon;Lee, Sung-Soo
    • Journal of Life Science
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    • v.20 no.3
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    • pp.381-387
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    • 2010
  • To find offspring of Jeju Black cattle (JBC) produced by embryo transfer (ET) and artificial insemination (AI), a molecular genetic study was carried out in candidate cattle populations collected from cattle farms in Jeju Island, Korea. The genetic marker set was composed of 11 ISAG microsatellite (MS) markers, 11 SAES MS markers selected by our preliminary analysis for population diversity of JBC and two major coat color related genes: MC1R and ASIP. The results showed a combined non-exclusion probability for first parent (NE-P1) that was higher than that recommended by ISAG (above 0.9995), and a combined non-exclusion probability for sib identity of $5.3{\times}10^{-10}$. Parentage analysis showed that the cases identified the candidate's father only (77.0%), mother only (54.0%), and both parents (40.5%) in the candidate offspring population. The ET and AI calves were identified as 14.7% in the in vitro fertilized eggs provided and 32.4% in total population, respectively. However, the result from ISAG marker analysis showed 3 identical allele-combinations in 7 calves, and that from ISAG/SAES MS marker combination also showed 1 identical allele-combination in 2 calves. Data from MS and coat-color gene analyses provided information for complete identification of all animals tested. Because the present JBC population was mostly bred using small nuclear founders through bioengineering techniques such as AI and ET, the genetic diversity levels obtained from MS analysis in the JBC population were relatively lower than those of other cattle populations, including Hanwoo. The results suggested that the more efficient marker combinations, including coat color related genotypes, should be studied and used for constructing a system for identification and molecular breeding of JBC as well.

A New Short Growth-Duration Rice Cultivar, "Keumo 3" (소득작물 전후작용 단기성 벼 품종 "금오3호")

  • Kang, Jong-Rae;Lee, Jong-Hee;Kwack, Do-Yeon;Lee, Jeom-Sik;Park, No-Bong;Ha, Woon-Gu;Park, Dong-Soo;Yeo, Un-Sang;Lim, Sang-Jong;Oh, Byeong-Geun
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.292-298
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    • 2009
  • A new rice cultivar "Keumo 3" was developed for adopting under double cropping system with after or before cash crop cultivation. It was selected from the cross-combination between YR17202 $F_2$/Shinkeumobyeo//YR15727-B-B-B-102. The parent, YR17202 $F_2$ individual plant, was used for tolerance to lodging, it derived from a cross between Nonganbyeo/Shinkeumobyeo. Nonganbyeo is well known to lodging tolerance cultivar, as well as biotic stress, because it was developed by crossing with Tongil type. And the YR15727-B-B-B-102 line used as another parent with short growth duration, likewise highly resistance to rice blast disease. The pedigree derived from the cross-combination YR17202 $F_2$/Shinkeumobyeo//YR15727-B-B-B-102 were generated to $F_7$, and a best line among them named as Milyang 201. After a series of yield trials, including local adaptability test conducted throughout the peninsular of Korea, Milyang 201 was registered with the name of "Keumo 3" in 2005. The cultivar belongs to a early maturing group and heads 4 days earlier than Keumobyeo, a standard cultivar. It has short culm, and less spikelet number per panicle than Keumobyeo. However, its milled rice yield grown under extremely late transplanting time, 10. July, over the 3 local sites for 2003-2005 years, averaged 4,48 MT/ha, which is 6% higher than the standard, Keumobyeo. "Keumo 3" has showed a durable resistance to leaf blast disease during fourteen blast nurseries screening covered from south to north in Korea for 2003-2007 years. And it was confirmed harbours pi-zt, a durable blast resistance gene. Moreover it was incompatible with 19 blast isolates under artificial inoculation, except one isolate, K1101. Additionally, "Keumo 3" exhibits resistance to $K_1$, $K_2$ and $K_3$ of bacterial blight pathogen, as well as strip virus disease resistance, and moderate resistance to dwarf virus disease. Consequently, the new rice cultivar "Keumo 3" would be well adopted where a bio stress makes a big problem annually.

The Effect of Photomodulation in Human Dermal Fibroblasts (피부 섬유아세포에서 광자극의 효과)

  • Kim, Mi Na;Kwak, Taek Jong;Kang, Nae Gyu;Lee, Sang Hwa;Park, Sun Gyoo;Lee, Cheon Koo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.4
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    • pp.325-331
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    • 2015
  • Skin is exposed to sunlight or artificial indoor light on a daily. The reached solar light on the earth surface consist of 50% visible light and 45% infrared (IR) except for ultra violet (UV). The negative effects of UV including UVB and UVA have been steadily investigated within the last decades. However, little is known about the effects of visible or IR light. In this study, we irradiated human dermal fibroblasts using light emitting diode (LED) to investigate the optimal parameter for enhancing cell growth and collagen synthesis. We found that red of 630 nm and green of 520 nm enhance the cell proliferation, but irradiation with purple and blue light exerts toxic effects. To examine the response of irradiation time and light intensity on the fibroblasts, cells were exposed to red or green light with intensities from 0.05 to $0.75mW/cm^2$. Procollagen secretion was increased of 1.4 fold by 10 min irradiation, while 30 min treatment decreased the collagen synthesis of dermal fibroblasts. Treatment with red of $0.3mW/cm^2$ and green of 0.15 and $0.3mW/cm^2$ resulted in enhancement of collagen mRNA. Lastly, we investigated the combinatorial effect of red and green light on dermal fibroblasts. The sequential irradiation of red and green light is an efficient way for the purpose of the increase in the number of fibroblasts than single light treatment. On the other hand, the exposure of red light alone was more effective method for enhancing of collagen secretion. Our study showed that specific light parameters accelerated cell proliferation, gene expression and collagen secretion on human dermal fibroblasts. In conclusion, we demonstrate that light exposure with specific parameter has beneficial effects on the function of dermal fibroblasts, and suggests the possibility of its cosmetically and clinical application.

Phylogeographic and population genetic study of a Korean endemic freshwater fish species, Zacco koreanus (한국 고유 담수어종 참갈겨니(Zacco koreanus) 개체군의 계통지리학 및 집단유전학 연구)

  • Kim, Yu Rim;Jang, Ji Eun;Choi, Hee-kyu;Lee, Hyuk Je
    • Korean Journal of Environmental Biology
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    • v.38 no.4
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    • pp.650-657
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    • 2020
  • We conducted a phylogeographic analysis of Korean endemic Zacco koreanus populations inhabiting the East-flowing river (Gangneung Yeongokcheon; GY, Yangyang Namdaecheon; YN), the Han River (Seomgang; SG, Soksacheon; SS), and the Nakdong River(Gilancheon; GA) using the mitochondrial DNA cytochrome oxidase I (COI) gene (619 bp). Population genetic analysis was further performed to assess the population connectivity for the GY river where there is a large number of human-made artificial weirs with several fishways. The phylogeographic analysis revealed that while the populations of the East-flowing river and those of the Han River formed a monophyletic lineage, the Nakdong River individuals represented a distinct lineage with 3.7-4.2% (mean=4.0%) genetic distance from the other lineages. The population genetic analysis of the GY showed that a mid-stream population harbored relatively higher mitochondrial diversity relative to up- and down-stream populations, and there was no genetic differentiation between these three populations. The latter findings might suggest high genetic connectivity between the populations via genetic flow along the fishways. However, an analysis using faster-evolving genetic markers, such as microsatellites, is needed to confirm the findings of high population connectivity. Our study suggests the possibility of the presence of cryptic species in Z. koreanus in the Nakdong River basin. However, further study with more individual samples as well as additional markers or even more advanced genomic tools is required to test our hypothesis. Ecological or phenotypic analyses should be conducted to test whether the observed Nakdong River lineage represents a different or cryptic species, or simply hidden, but excessive, intraspecific diversity.