• Proceedings of the KSAR Conference
• /
• 2004.06a
• /
• pp.237-237
• /
• 2004

Parthenogenesis and culture condition are essential to intracytoplasmic sperm injection and cloning by nuclear transfer. This study investigated apoptosis and in vitro development of parthenogenetic preimplantation porcine embryos. 42∼44 h in vitro matured oocytes derived from a local abattoir were used. Apoptotic cell death was analyzed by using a terminal deoxynucleatidyl transferase mediated deoxyuridine 5-triphoshate nick-end labling (TUNEL) assay. (omitted)

• Proceedings of the KSAR Conference
• /
• 2004.06a
• /
• pp.217-217
• /
• 2004

The present study was conducted to investigate the developmental competency between male- and female-somatic cell derived nuclear-transferred porcine embryos, and the productive and survival efficiency of cloned male and female piglets. The potential of eggs receiving somatic cells to develop into blastocysts was not different among donor cells of different origins. (omitted)

• Reproductive and Developmental Biology
• /
• v.34 no.1
• /
• pp.15-19
• /
• 2010

Oocyte enucleation is essential for somatic cell nuclear transfer (SCNT) in the production of cloned animals or embryonic stem cells from adult somatic cells. Most studies of oocyte enucleation have been performed using micromanipulator-based techniques, which are technically demanding, time-consuming, and expensive. Several recent studies have used chemical-induced oocyte enucleation; however, each has been plagued by low efficiency and toxicity. In this study, I found that the co-treatment of murine oocytes with demecolcine and BMI-1026, a potent cdk1 inhibitor, resulted in a high enucleation rate (97%). This method is entirely independent of a micromanipulator and is suitable for the large-scale production of enucleated oocytes. This new method of enucleation will be useful in SCNT and in the development of handmade cloning techniques.

• Journal of Photoscience
• /
• v.4 no.3
• /
• pp.141-145
• /
• 1997

Polymerase chain reaction(PCR) was conducted with a pea cDNA library using two primers synthesized from homology analysis of amino acid sequences for animal and plant cytosolic FBPases. A PCR product with 650 bp long was cloned into pGEM-T vector and sequenced. The deduced amino acid sequence of the cDNA fragment was 98, 91, and 85% homologous with those of cytosolic FBPases from spinach, sugarbeet, and sugarcane, respectively. It was 51% homologous with amino acid sequence of FBPase from pea chloroplasts. Northern blot analysis was proceeded with the cDNA clone resulting that 1.2 kb transcript was highly expressed in light-grown pea leaves but almost not expressed in dark-grown etiolated pea seedlings. When peas grown in the light for 10 days were transferred to darkness, the transcript was gradually decreased with dark treatment, indicating that the expression of the enzyme was induced by continuous white light but suppressed by dark treatment. Pea cytosolic FBPase was highly expressed in leaves with trace amounts in stems. but almost not expressed in roots.

• Journal of Embryo Transfer
• /
• v.9 no.2
• /
• pp.161-166
• /
• 1994

This experiment was carried out to produce cloned aniraals by nuclear transplantation in rabbits. The ovulated oocytes were collected from the oviducts between 14 and 15 hours after hGG injection. The denuded oocytes were used as nuclear recipient cytoplasm following enucleation by micromanipulation. The blastomeres separated from the 8-cell embryos were used as nuclear donor. The nucleated oocytes receiving a blastomere in the perivitelline space were electrically fused in the 0.28 M mannitol solution at 1.5 kV /cm, 60$\mu$sec for three times. The nuclear transplant embryos which were used and developed to 2- to 4-cell stage in vitro were transferred into the oviducts of synchronized recipient does. A total of 64 nuclear transplant embryos were transferred to 7 recipient does and produced three offspring(4.7%) from a foster mother 31 days after embryo transfer.

• Animal cells and systems
• /
• v.2 no.3
• /
• pp.371-375
• /
• 1998

Twenty-one monoclonal anti-DNA autoantilndies were produced by fusing spleen cells from an autoimmune MRL/lpr mouse with SP2/0 myeloma cells. Hybridomas generated by the fusions were chosen for cloning on the basis of DNA binding by supernatant antibody. Each monoclonal antibody was purified to homogeneity and analyzed for the heavy and light chain isotypes and the binding specificity for single-stranded DNA, double-stranded DNA, and RNA. Sequence specificities and isoelectric points of the antibodies were also examined. All of the antibodies were lgG and tended to bind to both single-stranded and double-stranded DNA with a preference for the double-stranded form. Some of them also bound to RNA. Isoelectric points of the antibodies were shown to be high. The antibodies described in this report have characteristics of pathogenic anti-DNA antibodies.

• Animal cells and systems
• /
• v.4 no.3
• /
• pp.257-261
• /
• 2000

To elucidate phylogenetic relationships among poecilostome families 18S rDNA sequence data were generated for seven poecilostome and one cyclopoid copopods by PCR cloning and sequencing techmiques. Phylogenetic trees were constructed by maximum parsimony, neighbor joining, and maximum likelihood methods using cyclopoid sequence as an outgroup. The results from three different analyses showed that the seven poecilostome families were eiridel into two groups: Clausidiidae-Myicolidae-Synaptiphillidae-bomolochidae and Lichomologidae-Chondracanthidae-Ergasilidae. The molecular phylogenies were consistent with those from the morphological characters. Therefore, these analyses porvide further evidence for the utility of 18S rDNA sequences in addressing phylogenetic relationships among poecilostome families.

• Journal of The Korean Association For Science Education
• /
• v.26 no.5
• /
• pp.646-659
• /
• 2006

This study explored, through informal, conversation-type interviews, how college students relate to science in general as well as to two specific socioscientific issues: human cloning and animal dissection/experimentation. How students "relate" includes what kinds of attitudes they have toward science and socioscientific issues, how seriously they consider and want to engage with these issues, and how they express their opinions or make a decision. The sample (16 college students) was heterogeneous in terms of academic background, ethnicity, and school year. Each interview lasted for about one hour with audiotaping. Results indicated that most participants immediately brought in their own values and feelings in implicit or explicit ways. However, the depth of their personal engagements varied. Most of the participants either did not take socioscientific issues seriously or merely quoted their own values in resignation, seemingly not able to deal with the issues and overwhelmed by many other aspects of the issues. By reflecting on the participants' reactions, the discussion addresses some of the larger issues for current secondary science teaching that involve raising responsible democratic citizens.

• Proceedings of the KSAR Conference
• /
• 2003.06a
• /
• pp.18-18
• /
• 2003

In human, sudden infant death syndrome(SIDS) is synonyms for the sudden, unexpected and unexplained death of an infant. The incidence of SIDS has been estimated to be from 1 to 3%. Cloning has a relatively high rate of late abortion and early postnatal death, particularly when somatic cells are used as donors of nuclei and rates as high as 40 to 70% have been reported. However, the mechanisms for SIDS in cloned animals are not known yet. To date, few reports provide detailed information regarding phenotypic abnormality of cloned pigs. In this study, most of the cloned piglets were alive at term and readily recovered respiration. However, approximately 82% of male cloned piglets (81/22) died within a week after birth. Significant findings from histological examinations showed that 42% of somatic cloned male piglets died earlier than somatic cloned female piglets, most probably due to severe congestion of lung and liver or neutrophilic inflammation in brain, which indicates that unexpected phenotypes can appear as a result of somatic cell cloning. No anatomical defects in cloned female piglets were detected, but three of the piglets had died by diarrhea due to bacterial infection within 15 days after birth. Although most of male cloned piglets can be born normal in terms of gross anatomy, they develop phenotypic anomalies that include leydig cell hypoplasia and growth retardation post-delivery under adverse fetal environment and depigmentation of hair- and skin-color form puberty onset. This may provide a mechanism for development of multiple organ system failure in some cloned piglets. Th birth weights of male cloned pig in comparison with those of female cloned piglets are significantly reduced(0.8 vs 1.4kg) and showed longer gestational day(120 vs 114). In conclusion, brain meningitis and hepatopneumonic congestion are a major risk factor for SIDS and such pregnancy in cloned animals requires close and intensive antenatal monitoring.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.8
• /
• pp.1180-1184
• /
• 2006

The capsule that surrounds Yersinia pestis cells is composed of a protein-polysacchride complex; the purified protein component is fraction I (F1) antigen. We report the cloning of the cafl gene and its expression in Escherichia coli using the vector pETl02/D-TOPO and the F1-specific monoclonal antibody. The recombinant F1 (rF1) antigen had a molecular size of 17.5 kDa, which was identical to that of the F1 antigen produced by Y. pestis. Recombinant F1 protein was found to react to polyclonal antiserum to Y. pestis Fl. Recombinant F1 was purified by ProBond purification system and induced a protective immune response in BALB/c mice challenged with up to 10$^5$ virulent Y. pestis. Purified rF1 protein was used in an ELISA to evaluate the ability of a method to detect antibodies to Y. pestis in animal sera. These results strongly indicated that the rF1 protein is a suitable species-specific immunodiagnostic antigen and vaccine candidate.