• Title/Summary/Keyword: rDNA sequence

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Sex Detection and In Vitro Development of Biopsied Bovine Embryo for LAMP Based Embryo Sexing (LAMP 방법에 의한 소 수정란의 성 판별과 Biopsy에 따른 수정란의 체외발달)

  • Cho S. R.;Choi S. H.;Kim H. J.;Han M. H.;Choe C. Y.;Chung Y. G.;Son D. S.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.169-176
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    • 2005
  • Loop-mediated isothermal amplification (LAMP) is novel DNA amplification methods that amplifies a target sequence specifically under isothemal condition. The present study was to assess the in vitro viability afier biopsy and sexing rate of different types of embryo biopsied. In vivo compact morulae and blastocyst embryos were obtained from Korean Native Cow (KNC) superovulated with FSH (Antorin, R-10) on 7 Day after artificial insemination. in vitro compact morulae and blastocyst embryos were obtained with KNC or Holsteins that were gained on 6, 7 or 8 day after in vitro fertilization(IVF) with frozen semen. Biopsy of bovine embryo was carried out in a $80{\mu}l$ drop with $Ca^{2+}-Mg^{2+}$ free D-PBS and the viability of biopsied embryos were evaluated in IVMD (IFP, Japan) medium at 12 hrs culture time. The sex ratio of biopsied Hanwoo embryos were male vs. female of $43.5\%\;vs.\;56.5\%$ in vivo and $33.9\%\;vs.\;49.2\%$ in vitro respectively, and male rate of biopsied Holstein embryos were significantly higher than female $(70.8\;vs.\;29.2\%)$. and indefinite rate of in vitro embryos was $16.9\%$ and in vivo was not. The degeneration rate of biopsied embryo, in vitro embryos were significantly higher than in vivo $(13.2\%\;vs,\;0.0\%,\;p<0.05)$. The survivability of in vivo embryo were between biopsied following punching method was significantly (P<0.05) higher than bisection method produced embryos $(100\%\;vs.\;83.3\%)$ and in vitro had no difference. However, the degeneration rate of biopsied embryo by bisection method was significantly higher than punching methods between in vivo and in vitro $(16.7\;vs.\;22.6\%,\;respectively,\;p<0.05)$. In conclusion, these results indicate that punching method was optimal and survivability after embryo biopsy was useful for reducing the damage caused by the embryo biopsy procedure for LAMP-based embryo sexing.

Physiological and Biochemical Characterization of Bacillus spp. from Polychaete, Perinereis aibuhitensis (갯지렁이(Perinereis aibuhitensis)에서 분리한 Bacillus spp.의 생리생화학적 특성 분석)

  • Shin, Seyeon;Yundendorj, Khorloo;Lee, Sang-Suk;Kang, Kyoung-Ho;Kahng, Hyung-Yeel
    • Journal of Life Science
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    • v.23 no.3
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    • pp.415-425
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    • 2013
  • This study compared the characteristics of five Bacillus strains capable of aerobic and anaerobic growth, CBW3, CBW4, CBW9, CBW14 and EBW10. They were isolated and selected from a polychaete, Perinereis aibuhitensis, which is known as a good degrader of organic compounds in marine wetland. Based on a 16S rRNA sequence, CBW3 and CBW14 were found to share more than 99.8% similarity with B. nanhaiensis, B. arsenicus and B. barbaricus. CBW4, CBW9 and EBW10 shared 92.7%, 99.8%, and 99.8% similarity with B. anthracis, B. algicoa and B. thuringiensis, respectively. The temperature, salinity, and pH ranges of the cell growth of the Bacillus strains were $4-45^{\circ}C$, 0-17%, and pH 5-pH 9, respectively. All Bacillus strains were found to exhibit enzyme activities for the degradation of casein and starch. Notably, strain EBW10 exhibited the enzyme activities for all the tested macromolecules, DNA, casein, starch, cellulose, and four kinds of Tweens, which suggests the possibility that it had protease, amylase, cellulose, and lipase. All five Bacillus strains had alkaline phosphatase activities, and the strains CBW3, CBW4, and EBW10 also had acid phospatase. Strains CBW3 and EBW10 exhibited the enzyme activities both for esterase (C4) and esterase lipase (C8). The analysis of fatty acids revealed that in all strains, major fatty acids were anteiso $C_{15:0}$ and iso $C_{15:0}$.

Quality Evaluation by the Addition of Pine Needle and Artemisia princeps Extracts in Vinegared Kochujang (솔잎, 약쑥추출물을 첨가한 초고추장의 품질평가)

  • Kim, Eun-Lyang;Kang, Sun-Chul
    • Applied Biological Chemistry
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    • v.50 no.3
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    • pp.167-177
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    • 2007
  • This study was carried out to develop natural vinegared kochujang. We first formulated vinegared kochujang containing pine needle (PN) and Artemisia princeps (AP) extracts. A dominant strain was isolated from the vinegared kochujang and identified as Bacillus amyloliquefaciens ER282 based on the 16S rDNA sequence. The plant extracts of PN and AP were also investigated for their antibacterial activity and showed enough good activities to use for vinegared kochujang as a natural preservative. Quality characteristics of vinegared kochujang were evaluated, based on total cell numbers, pH, acidity, sugar content, color and sensory evaluation during storage at $25^{\circ}C$ and $37^{\circ}C$, respectively, for 3 weeks. No significant differences were found in the pHs and acidities of vinegared kochujang added with the plant extracts of PN and AP during their storage compared to starting time; however, total cell numbers were gradually decreased during the storage at $25^{\circ}C$ and $37^{\circ}C$, as the storage period was increased. For sensory evaluation, overall preference continuously declined depending on storage period, but the addition of 3% extracts of PN could delay the quality loss of vinegared kochujang during storage at $25^{\circ}C$ and $37^{\circ}C$ for 3 weeks.

Genetic Variations of Chicken TYR Gene and Associations with Feather Color of Korean Native Chicken (KNC) (한국 토종닭 모색 변이와 TYR 유전자형 간의 상관관계 분석)

  • Choi, Jin Ae;Lee, Jun-Heon;Jang, Hyun-Jun;Lee, Kyung-Tai;Kim, Tae-Hun;Lee, Hyun-Jeong;Heo, Kang-Nyeong;Kim, Chong-Dae;Han, Jae-Yong;Park, Mi Na
    • Korean Journal of Poultry Science
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    • v.41 no.1
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    • pp.7-14
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    • 2014
  • Tyrosinase (TYR) gene is located on chromosome 1 in chicken and it is composed of five exons and four introns. TYR gene is described as a key enzyme in melanin biosynthesis. Most examples of complete albinism in chicken have been due to defects in the tyrosinase gene. The association of feather color and sequence polymorphism in the Tyrosinase (TYR) gene was investigated using Korean Native chicken H breed (H_PL), Korean Native chicken L/W breed(L/W_PL) and 'Woorimatdag' commercial chickens (Woorimatdag_CC). From L_PL and W_PL breed analyses, 4 synonymous SNPs (locus G33A, G116A, C217T and C247T) and 2 SNPs (G838A and G958A) were detected in 4th exon and 4th intron of TYR gene respectively. The genotype frequencies for 6 SNPs were compared between L_PL and W_PL and W_PL represented homozygous SNP types in all the analyzed SNP positions while L_PL displayed various SNP types.