• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2003.10a
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• pp.121-121
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• 2003

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.4
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• pp.482-486
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• 2007

Basic objective of this research was to compare the milk yield and composition of New Zealand White transgenic rabbit females expressing recombinant human factor VIII (hFVIII) in mammary gland during lactation with that of non-transgenic rabbit females of the same age during 30 days of lactation. Transgenic founders were generated by the microinjection of foreign DNA (mWAP-hFVIII gene construct) into the egg. F1, F2 and F3 generations of transgenic rabbits were obtained after mating of transgenic founder rabbits with non-transgenic rabbits. The amount of milk rejected was measured by weight-suckle-weight method at $10^{th}$, $20^{th}$and $30^{th}$ day of lactation. Quality of milk (content of fat, protein, lactose, dry ash, and some minerals) from transgenic and non-transgenic rabbit was also determined. Comparison of milk yield, determined by weight-suckle-weight method, showed significantly higher (p<0.05) milk production at day 20 of first lactation in non-transgenic females, but on the same day of second lactation higher milk yield was measured in transgenic ones. Significantly higher (p<0.05) content of milk fat and protein was determined in transgenic milk whilst higher content of lactose was found in non-transgenic milk. The content of minerals (calcium, phosphorus, magnesium and sodium) did not differ in both experimental and control groups. Our results showed that milk yield and composition of transgenic rabbit females (mammary specific transgenic over-expression of hFVIII) over several generations is only slightly and transiently different from milk yield of non-transgenic females, which had no significant consequence on the litter size and viability.

• Korean Journal of Animal Reproduction
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• v.17 no.4
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• pp.375-383
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• 1994

The human growth hormone (hGH) gene uder the control of the rat $\beta$-casein promoter gene was designed to produce transgenic mouse expressed hGH gene in only mammary gland. One hundred seventy two eggs microinjected were transferred to the oviducts of pseudopregnants and 43 offspring were delivered. By Southern blotting hybridization, 3 were transgenic with rat $\beta$-casein/hGH gene. The copy numbers of three transgenic founder were 1, 5, and 15, respectively. A radioimmunoassay was developed to quantitate the amount of expression of the hGH gene in mammary gland of transgenic mice. The amount of hGH was 13.3ng/ml in the lactating milk of one transgenic line, showing predominantly higher than 3.0ng/ml in milk of control mice. Therefore, our findings suggested that $\beta$-casein promoter may induce the tissue specific expression of structural gene.

• Korean Journal of Animal Reproduction
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• v.16 no.3
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• pp.225-230
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• 1992

Many transgenic mice expressing human growth hormone gene were infertile. To investigate the infertility of these transfenic mice, it was looked into the estrus cycle and sexual behaviour and also tested through in vitro fertilization whether the germ cells of these mice normal or not. The infertile female transgenic mice were mated to the fertile males of ICR strain, but in almost all of them the vaginal plugs were not detected and their estrus cycles by vaginal smear were almost irregular which kept up estrus or diestrus stage. Many male transgenic mice did not have the ability of sexual behaviour. Therefore the viability of germ cells in infertile male transgenic mice was investigated by in vitro fertilization, but the sperm were normally fertilized with the eggs and the transgene of parent was passed on to the progeny. These results consequently suggest that the infertility of transgenic mice experssing human growth hormone gene may be due to the physiological activity of human growth hormone, not germ cells.

• Korean Journal of Animal Reproduction
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• v.14 no.3
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• pp.191-197
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• 1990

The transgenic mice were produced by microinjection of human growth hormone gene fused with mouse metallothionein Ⅰ promoter. They were mated with momal mice by backcross or brother-sister mating. The reproduction efficiencies of female and male n the FO transgenic mice were 17.6%(3/17 mice) and 31.2%(5/16 mice), respectively, and were very lower than that in normal mice(85.7% and 100%, respectively). Interestingly, a few of female transgenic mice were fertile which was different from the previous reports. Of 6 fertile transgenic mice, 2 mice were identified as mosaic type by the reduced frequency of genetic transmission to successive generation below Mendelian levle and the enhanced copy numbers of transgene in progeny mice compared with the transgenic parent. In the group of F1, F2, F3 transgenic mice the reproduction efficiencies of males were gradually improved, whereas females were absolutely infertile. It was consequently shown that the transgenic mice expressing human growth hormone gene were frequently infertile, but the genotypic and phenotypic characteristics of the fertile transgenic mice were normally passed on to the progeny through herm line. Therefore it must be considered wheter or not the products of foreign DNA introduced into animals will detrimentally affect their physiological aspects.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.190-190
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• 2004

Tissue plasminogen activator (tPA) plays important roles in the brain after excitotoxic injury. This study was conducted to produce transgenic pig harboring human tissue plasminogene activator (htPA) gene. Recombinent htPA(rhtPA) genes containing bovine-β-casein promoter (bBC) were prepared for microinjection and testified the expression level of htPA protein from the Chinese hamster ovary (CHO) cell lines before NDA microinjection into the porcine pronuclei. (omitted)

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1369-1373
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• 2004

The percentages of sperm motility and normal acrosome on the liquid boar semen diluted and preserved at $4^{\circ}C$ with lactose hydrate, egg yolk and N-acetyl-D-glucosamine (LEN) diluent were significant differences according to preservation day and incubation time, respectively. The sperm motility steadily declined from 96.9% at 0.5 h incubation to 78.8% at 6 h incubation at 1 day of preservation. However, the sperm motility rapidly declined after 4 day of preservation during incubation. The normal acrosome steadily declined from 93.3% at 0.5 h incubation to 73.8% at 6 h incubation at 1 day of preservation. However, the normal acrosome rapidly declined after 3 day of preservation during incubation. The rates of sperm penetration and polyspermy were higher in 5 and $10{\times}10^6$ sperm/ml than in 0.2 and $1{\times}10^6$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in $10{\times}10^6$ sperm/ml compared with other sperm concentrations. The rates of blastocysts from the cleaved oocytes (2-4 cell stage) were highest in $1{\times}10^6$sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at $4^{\circ}C$ could be used for in vitro fertilization of pig oocytes matured in vitro. Also, we recommend $1{\times}10^6$sperm/ml concentration for in vitro fertilization of pig oocytes.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.6
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• pp.789-793
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• 2012

Fertilization of the oocyte commences embryogenesis during which maternally inherited mRNAs are degraded and the embryonic genome is activated. Transcription of embryonic mRNA is initiated by embryonic genome activation (EGA). RNA polymerase II (RNA Pol II) is responsible for the synthesis of mRNAs and most small nuclear RNAs, and consists of 12 subunits, the largest of which characteristically harbors a unique C-terminal domain (CTD). Transcriptional activity of RNA Pol II is highly regulated, in particular, by phosphorylation of serine residues in the CTD. Here, we have shown the presence of RNA Pol II CTD phosphoisoforms in porcine oocytes and preimplantation embryos. The distribution pattern as well as phosphorylation dynamics in germinal vesicles and during embryogenesis differed in developmental stages with these isoforms, indicating a role of RNA Pol II CTD phosphorylation at the serine residue in transcriptional activation during both oocyte growth and embryonic genome activation. We additionally examined the effects of the RNA Pol II inhibitor, ${\alpha}$-amanitin, on embryo development. Our results show that inhibition of polymerase, even at very early stages and for a short period of time, dramatically impaired blastocyst formation. These findings collectively suggest that the functionality of maternal RNA Pol II, and consequently, expression of early genes regulated by this enzyme are essential for proper embryo development.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.3
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• pp.307-312
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• 2007

The present study was carried out to investigate differentially expressed chicken muscle proteins using proteomics approach. More than 300 protein spots were investigated for the muscle samples in 2DE gels and the differentially expressed protein spots between pectoralis and peroneus longus muscles from Cornish and White Leghorn breeds were characterized by MALDI-TOF. In pectoralis muscles, PGAM1 protein was detected as differentially expressed between White Leghorn and Cornish breeds. On the other hand, 4 protein spots (SP22, nxf-2, SOD1, TNNI2) were differentially expressed between White Leghorn and Cornish breeds in peroneus longus muscles. These proteins assumed to be related with muscle development, growth, stress, and movements in chicken. In this experimental process, 2D reference map of the chicken muscle proteins was needed and 25 proteins, which were commonly expressed in both pectoralis and peroneus longus muscles in both breeds, were selected and characterized. Upon finishing the exact roles of the differentially expressed proteins, the identified 5 proteins will be used as valuable information for the fundamental mechanisms of muscle biology and underline genetics.

• BMB Reports
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• v.31 no.4
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• pp.413-417
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• 1998

Transgenic mice containing a bovine ${\beta}-Casein/Human$ lysozyme fusion gene (pBZ) were generated in order to produce human lysozyme in their milk. The expression vector was a quadripartite fusion consisting of a 2 kb upstream DNA of the bovine ${\beta}-casein$ gene, human lysozyme gene, intron II of the rabbit ${\beta}-globin$ gene, and the polyadenylation/termination signals of SV40 DNA. Fertilized mouse zygotes were microinjected with pBZ, then transferred into the oviduct of foster mothers. Out of 20 mice born, 11 survived until postweaning and three were identified as positivetransgenic by Southern blot analysis (one male and two females). The founder mice were mated to BCFl mice to produce transgenic progeny. It was confirmed by RT-PCR and Northern blot analyses that the transgene was specifically expressed in the mammary gland of the founder mice. Furthermore, the artificial introns within the transgenic RNA was proven to be correctly spliced out as judged by RT-PCR analysis. These results indicated that transgenic mice generated in this study properly expressed the human lysozyme RNA in their mammary gland.