Rat H - Y 항체에 의한 생쥐 Embryo 의 성 조절에 관한 연구 Ⅲ. H - Y 항혈청에 의한 BALB c 생쥐 상실배의 성 판별

정장용(J . Y . Jeung),박충생(C . S . Park),박희성(H . S . Park) 한국축산학회 1989 한국축산학회지 Vol.31 No.10

This experiment was carried out to develop a new technique by identifying XX-bearing embryos prior to implantation by immunological method. H-Y antiserum was prepared in inbred Wistar female rats by repeated immunization with spleen cells from males of the same strain. The reactivity of H-Y antibody was confirmed by culturing mouse embryos in the medium containing H-Y antiserum and complement obtained from the guinea pig. The optimal condition for the activity of H-Y antibody was also investigated by culturing embryos under the various conditions of equilibration times, complement concentrations and various media. The results obtained in this experiment are as follows: When the embryos were cultured in the medium of H-Y antiserum and complement which was given the equilibration time of less than 30 minutes in CO₂ incubator, the lysis-rate of embryo was 89.3%. The embryo lysis-rates in the equilibration time of 1-1.5, 3-3.5, or 24-26 hours were 48.1, 47.7 and 48.2%, respectively. When the concentration rate of complement to H-Y antiserum varied from 0.25 - 4.0, the lysis-rate of embryo was 43.2 to 52.7%. The concentration rate of complement did not influence the lysis-rate of embryos. The meda of D-PBS + 0.3% BSA, D-PBS + 20% FCS, Ham`s F-10 + 0.3% BSA and Ham`s F-10 + 20% FCS showed the embryo lysis-rate of 46.4, 57.4, 49.3 and 49.1%, respectively. The culture media used in this experiment did not show any significant difference in the embryo lysis-rate. After the embryos were cultured to the late blastocyst in the media of D-PBS + NGPS + H-Y antiserum or D-PBS + NGPS + normal female rat serum the normally developed embryos were selected and transferred to the pseudo pregnant recipients. The percentages of their female offspring were 82.3%(14/17) in H-Y antiserum treatment and 53.6%(15/28) in normal serum treatment and showed a significant difference between the two treatments(p $lt;0.001).

Rat H-Y 항체에 의한 생쥐 Embryo 의 성 조절에 관한 연구 1 . H-Y 항체의 처리가 생쥐 상실배 (桑實胚) 의 발달에 미치는 영향 및 세포독성시험

정장용(J . Y . Jeung),박충생(C . S . Park),박희성(H . S . Park) 한국축산학회 1989 한국축산학회지 Vol.31 No.8

This experiment was carried out to develop a new technique by immunological method. H-Y antiserum was prepared in inbred Wistar female rats by repeated immunization with new-born testis supernatant and spleen cells from males of the same strain. The activity of H-Y antibody in antiserum was tested by cytotoxicity and biological tests. The results obtained in this experiment. are as follows: In the sperm cytotoxicity test it was showed about 70% of the sperm were dead in the 1/2 to 1/8 dilution of H-Y antiserum immunized with spleen cells, new-born testis or the compound of both antigens. As the dilution increased, the death rate of sperm decreased markedly. H-Y antibody absorbed with female-rat spleen cells showed higher death-rate of sperm than that with male-rat spleen cells. The normal female-rat serum showed the sperm death-rate of 14.6 to 27.9% irrespective of the dilution rate. The difference between the two sperm death-rates was significant. The embryos cultured in the medium of complement and H-Y antiserum immunized with male-rat spleen, new-born testis or the compound of both antigens showed the lysis-rates of 48.9, 50.0 and 46.3% respectively. There was no significant difference among each lysis-rate. But the lysis-rate of the embryos cultured in the medium of complement and normal female rat serum was 5.1% and it was markedly different from the above lysis rates(p $lt;0.001).

Effects of Y₂O₃ additions on the oxygen diffusion in top-seeded melt growth processed YBa₂Cu₃O_7-y superconductors

Jun, B.H.,Jung, S.A.,Park, S.D.,Park, B.J.,Han, Y.H.,Kim, C.J. North-Holland 2011 Physica. C, Superconductivity Vol.471 No.21

To understand the effect of Y<SUB>2</SUB>BaCuO<SUB>5</SUB> (Y211)/YBa<SUB>2</SUB>Cu<SUB>3</SUB>O<SUB>7-y</SUB> (Y123) interfaces on the oxygen diffusion in single grain YBa<SUB>2</SUB>Cu<SUB>3</SUB>O<SUB>7-y</SUB> superconductors, single grain Y123 superconductors with 0.05 and 0.3moles of Y<SUB>2</SUB>O<SUB>3</SUB> additions were fabricated by a top-seeded melt growth (TSMG) process. Y123 compacts with Y<SUB>2</SUB>O<SUB>3</SUB> additions were subjected to melt growth heating cycles with a cooling rate of 1<SUP>o</SUP>C/h through a peritectic temperature (1015<SUP>o</SUP>C) and then annealed at 450<SUP>o</SUP>C for 200h in flowing oxygen. The superconducting temperature (T<SUB>c</SUB>) and critical current density (J<SUB>c</SUB>) were estimated for the three different regions (top surface (s), intermediate (i) and center (c)) of samples. The amount of Y211/Y123 interface area in single grain Y123 superconductors was successfully controlled by Y<SUB>2</SUB>O<SUB>3</SUB> additions. The T<SUB>c</SUB> values of s regions were higher than those of i and c regions, which indicates the presence of more oxygen at the sample surfaces. In addition, the T<SUB>c</SUB> values of i and c regions of the Y123 sample with 0.3mole Y<SUB>2</SUB>O<SUB>3</SUB> addition were higher than those of the same regions of the Y123 sample with 0.05mole Y<SUB>2</SUB>O<SUB>3</SUB> addition due to the promoted oxygen diffusion through Y211/Y123 interfaces and other related defects. In spite of the promoted oxygen diffusion by Y<SUB>2</SUB>O<SUB>3</SUB> addition, the large T<SUB>c</SUB> difference among the regions still existed, which suggests sluggish oxygen diffusion into single Y123 grains.

Experimental infection of mandarin duck with highly pathogenic avian influenza A (H5N8 and H5N1) viruses

Kang, H.M.,Lee, E.K.,Song, B.M.,Heo, G.B.,Jung, J.,Jang, I.,Bae, Y.C.,Jung, S.C.,Lee, Y.J. Elsevier Scientific Pub. Co 2017 Veterinary microbiology Vol.198 No.-

<P>A highly pathogenic avian influenza (HPAI) H5N8 virus was first detected in poultry and wild birds in South Korea in January 2014. Here, we determined the pathogenicity and transmissibility of three different clades of 1-15 viruses in mandarin ducks to examine the potential for wild bird infection. H5N8 (Glade 2.3.4.4) replicated more efficiently in the upper and lower respiratory tract of mandarin ducks than two previously identified H5N1 virus clades (clades 2.2 and 2.3.2.1). However, none of the mandarin ducks infected with H5N8 and H5N1 viruses showed severe clinical signs or mortality, and gross lesions were only observed in a few tissues. Viral replication and shedding were greater in H5N8-infected ducks than in H5N1-infected ducks. Recovery of all viruses from control duck in contact with infected ducks indicated that the highly pathogenic H5 viruses spread horizontally through contact. Taken together, these results suggest that H5N8 viruses spread efficiently in mandarin ducks. Further studies of pathogenicity in wild birds are required to examine possible long-distance dissemination via migration routes. (C) 2016 Elsevier B.V. All rights reserved.</P>

Expression and N-glycan analysis of human 90K glycoprotein in Drosophila S2 cells

Kim, K.R.,Kim, Y.K.,Cheong, H.,Kim, J.Y.H.,Cha, H.J. IPC Science and Technology Press ; Elsevier Scienc 2013 Enzyme and microbial technology Vol.53 No.3

Human 90K (h90K; Mac-2-binding protein) glycoprotein is a potential pharmaceutical due to its inhibitory activity against cancer metastasis and expansion. Here, h90K glycoprotein was produced in insect Drosophila S2 cell system, and its N-glycan pattern was analyzed. A plasmid encoding h90K gene, fused with a hexahistidine tag under the control of Drosophila metallotionein promoter, was stably transfected into S2 cells. After copper sulfate induction, transfected S2 cells secreted recombinant h90K at a good expression level of 28mg/L in a 150-mL spinner flask culture. The purified recombinant h90K showed an apparent molecular weight of ~78kDa which was much smaller than that (~97kDa) of the natural h90K. Because de-N-glycosylated h90K appeared at ~60kDa protein band, it was suggested that the recombinant h90K from S2 cells has small N-glycans with about half the molecular weight (~18kDa) of N-glycans of the natural h90K. Through detail analyses using high-performance liquid chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, the S2-derived recombinant h90K was confirmed that it has simple paucimannosidic structures containing two or three mannose residues with core fucose as the major (~79%) N-glycans.

Haplotype and mutation analysis for newly suggested Y-STRs in Korean father-son pairs

Oh, Y.N.,Lee, H.Y.,Lee, E.Y.,Kim, E.H.,Yang, W.I.,Shin, K.J. Elsevier Science 2015 FORENSIC SCIENCE INTERNATIONAL GENETICS Vol.15 No.-

In this study, 363 Korean father-son haplotype transfers in 351 families were analyzed using an in-house multiplex PCR system for 14 Y-STRs (DYS385a/b, DYF387S1, DYS391, DYS449, DYS460, DYS481, DYS518, DYS533, DYS549, DYS570, DYS576, DYS627 and DYS643), that included 11 loci newly added to the PowerPlex Y23 system or the Yfiler Plus system. The Y-STRs showed gene diversity values ranging from 0.2499 to 0.9612; the multicopy Y-STR loci DYS385 and DYF387S1 had high gene diversity of 0.9612 and 0.9457, respectively. In addition, DYF387S1, which has two copies, showed three alleles in seven individuals, and micro-variant alleles were observed in 14 individuals at four loci (DYS448, DYS518, DYS570 and DYS627). Among 351 haplotypes for the 11 newly added Y-STRs, 350 different haplotypes were observed, with an overall haplotype diversity of 0.9999 and discrimination capacity of 99.72%. In 363 haplotype transfers from 351 pedigrees, 29 single-step mutations were observed at 11 Y-STRs. Locus-specific mutation rate estimates varied from 0.0 to 1.93x10<SUP>-2</SUP>, with an average estimated mutation rate of 6.66x10<SUP>-3</SUP>. Two father-son pairs had mutations at two different loci in 11 Y-STRs. The number of pairs with mutations at multiple loci increased to five when the mutation event was investigated for haplotype transfer at 28 Y-STRs including 17 Yfiler loci and 11 Y-STRs examined in this study: four father-son pairs had mutations at two loci, and one pair had mutations at three loci. Overall, mutations were frequently observed at DYS449, DYS576 and DYS627 loci, which are known to be rapidly mutating Y-STRs. Mutation rate estimates at most loci were not significantly different from rates in other populations, but estimates for DYF387S1, DYS518 and DYS570 were considerably lower in the Korean population than in other populations.

Selection for Duration of Fertility and Mule Duck White Plumage Colour in a Synthetic Strain of Ducks (Anas platyrhynchos)

H.C. Liu,J.F. Huang,S.R. Lee,H.L. Liu,C.H. Hsieh,C.W. Huang,M.C. Huang,C. Tai,J.P. Poivey,R. Rouvier,Y.S. Cheng 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.5

A synthetic strain of ducks (Anas platyrhynchos) was developed by introducing genes for long duration of fertility to be used as mother of mule ducklings and a seven-generation selection experiment was conducted to increase the number of fertile eggs after a single artificial insemination (AI) with pooled Muscovy semen. Reciprocal crossbreeding between Brown Tsaiya LRI-2 (with long duration of fertility) and Pekin L-201 (with white plumage mule ducklings) ducks produced the G0. Then G1 were intercrossed to produce G2 and so on for the following generations. Each female duck was inseminated 3 times, at 26, 29, and 32 weeks of age. The eggs were collected for 14 days from day 2 after AI. Individual data regarding the number of incubated eggs (Ie), the number of fertile eggs at candling at day 7 of incubation (F), the total number of dead embryos (M), the maximum duration of fertility (Dm) and the number of hatched mule ducklings (H) with plumage colour were recorded. The selection criterion was the breeding values of the best linear unbiased prediction animal model for F. The results show high percentage of exhibited heterosis in G2 for traits to improve (19.1% for F and 12.9% for H); F with a value of 5.92 (vs 3.74 in the Pekin L-201) was improved in the G2. Heritabilities were found to be low for Ie (h2 = 0.07±0.03) and M (h2 = 0.07±0.01), moderately low for Dm (h2 = 0.13±0.02), of medium values for H (h2 = 0.20±0.03) and F (h2 = 0.23±0.03). High and favourable genetic correlations existed between F and Dm (rg = 0.93), between F and H (rg = 0.97) and between Dm and H (rg = 0.90). The selection experiment showed a positive trend for phenotypic values of F (6.38 fertile eggs in G10 of synthetic strain vs 5.59 eggs in G4, and 3.74 eggs in Pekin L-201), with correlated response for increasing H (5.73 ducklings in G10 vs 4.86 in G4, and 3.09 ducklings in Pekin L-201) and maximum duration of the fertile period without increasing the embryo mortality rate. The average predicted genetic response for F was 40% of genetic standard deviation per generation of selection. The mule ducklings’ feather colour also was improved. It was concluded that this study provided results for a better understanding of the genetics of the duration of fertility traits in the common female duck bred for mule and that the selection of a synthetic strain was effective method of improvement.

돼지 H-FABP 유전자의 다형성 및 경제 형질과의 연관성 구명

최봉환,김태헌,이지웅,조용민,이혜영,조병욱,정일정 한국동물자원과학회 2003 한국축산학회지 Vol.45 No.5

The purpose of this study was to detect association between genetic variation and economic trait in the porcine heart type fatty acid-binding protein gene as a candidate gene for the traits related with growth and meat quality in pigs. The H-FABP is a 15-kDa protein expressed in several tissues with high demand for fat metabolism such as cardiac and skeletal muscle and lactating mammary gland. H-FABP is small intracellular protein involved in fatty acid transport from the plasma membrane to the site of β-oxidation and/or triacylglycerol or phospholipid synthesis. In this study, H-FABP PCR-RFLP was performed in F_(2) population composed of 214 individuals form an intercross between Korean Native Boars and Landrace sows. PCR products form tow primer sets within H-FABP gene were amplified in 850bp and 700bp. Digestion of PCR products with the restriction digestion enzymes HaeⅢ and Hinf Ⅰ, revealed fragment length polymorphisms(RFL. Ps). The genotype frequencies from H-FABP/HaeⅢ was .29 for genotype DD, .53 for genotype Dd, and .15 for genotype dd, respectively. The genotype frequencies of HH, Hh, and hh from H-FABP(hinf Ⅰ was .38, .41, and .20, respectively, in the population.Relationships between their genotypes and economic traits were estimated. In H-FABP/HaeⅢ locus, there were specific genotypes(Dd and dd) associated with economic traits such as body weight. In H-FABP/Hinf Ⅰ Iocus, Genotypes of HH and Hh associated with growth traits such as body weights at 5, 12, and 30 week of age (p<.05 or p<.001) and back fat thickness, body fat including abdominal and trimmed fat (p<.001) and intramuscular fat(p<.05). The 'H'allele was positivecly associated with gaining of body weight and fatness deposition. In conclusion, a significant association of the H-FABP gene from its genetic variation was found on body weight, intramuscular fat and backfat thickness.

Oxygen-dependent enhancement of hydrogen production by engineering bacterial hemoglobin in Escherichia coli

Jo, B.H.,Kim, J.Y.H.,Seo, J.H.,Cha, H.J. Pergamon Press ; Elsevier Science Ltd 2014 International journal of hydrogen energy Vol.39 No.20

H<SUB>2</SUB> production under aerobic conditions has been proposed as an alternative method to overcome the fundamentally low yield of H<SUB>2</SUB> production by fermentative bacteria by maximizing the number of electrons that are available for H<SUB>2</SUB>. Here, we engineered Vitreoscilla hemoglobin (VHb) in Escherichia coli to study the effects of this versatile oxygen (O<SUB>2</SUB>)-binding protein on oxic H<SUB>2</SUB> production in a closed batch system that was supplemented with glucose. The H<SUB>2</SUB> yields that were obtained with the VHb-expressing E. coli were greatly enhanced in comparison to the negative control cells in culture that started with high O<SUB>2</SUB> tensions. The formate hydrogen lyase (FHL) activity of oxically cultured, VHb-expressing cells was also much higher than that of the negative control cells. Through inhibitor studies and time-course experiments, VHb was shown to contribute to the improved H<SUB>2</SUB> yield primarily by increasing the efficiency of cellular metabolism during the aerobic phase before the onset of H<SUB>2</SUB> production and not by working as an O<SUB>2</SUB>-scavenger during H<SUB>2</SUB> production. This new approach allowed more substrate to remain to be further utilized for the production of more H<SUB>2</SUB> from limited resources. We expect that VHb can be successfully engineered in potential aerobic H<SUB>2</SUB>-producing microbial systems to enhance the overall H<SUB>2</SUB> production yield. In addition, the remarkably high FHL activity of oxically grown, VHb-expressing cells may make this engineered strain an attractive whole-cell biocatalyst for converting formate to H<SUB>2</SUB>.

Investigation into the sequence structure of 23 Y chromosomal STR loci using massively parallel sequencing

Kwon, S.Y.,Lee, H.Y.,Kim, E.H.,Lee, E.Y.,Shin, K.J. Elsevier Science 2016 FORENSIC SCIENCE INTERNATIONAL GENETICS Vol.25 No.-

Next-generation sequencing (NGS) can produce massively parallel sequencing (MPS) data for many targeted regions with a high depth of coverage, suggesting its successful application to the amplicons of forensic genetic markers. In the present study, we evaluated the practical utility of MPS in Y-chromosome short tandem repeat (Y-STR) analysis using a multiplex polymerase chain reaction (PCR) system. The multiplex PCR system simultaneously amplified 24 Y-chromosomal markers, including the PowerPlex<SUP>®</SUP> Y23 loci (DYS19, DYS385ab, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS481, DYS533, DYS549, DYS570, DYS576, DYS635, DYS643, and YGATAH4) and the M175 marker with the small-sized amplicons ranging from 85 to 253bp. The barcoded libraries for the amplicons of the 24 Y-chromosomal markers were produced using a simplified PCR-based library preparation method and successfully sequenced using MPS on a MiSeq<SUP>®</SUP> System with samples from 250 unrelated Korean males. The genotyping concordance between MPS and the capillary electrophoresis (CE) method, as well as the sequence structure of the 23 Y-STRs, were investigated. Three samples exhibited discordance between the MPS and CE results at DYS385, DYS439, and DYS576. There were 12 Y-STR loci that showed sequence variations in the alleles by a fragment size determination, and the most varied alleles occurred in DYS389II with a different sequence structure in the repeat region. The largest increase in gene diversity between the CE and MPS results was in DYS437 at +34.41%. Single nucleotide polymorphisms (SNPs), insertions, and deletions (indels) were observed in the flanking regions of DYS481, DYS576, and DYS385, respectively. Stutter and noise ratios of the 23 Y-STRs using the developed MPS system were also investigated. Based on these results, the MPS analysis system used in this study could facilitate the investigation into the sequences of the 23 Y-STRs in forensic genetics laboratories.