The purpose of this study was undertaken to evaluate of cryopreservation efficiency in α 1,3-galactosyltransferase knock-out(GalT KO) cloned miniature pig sperm. To compare ability of frozen-thawed sperm characteristics, three different pig strains (GalT KO) cloned miniature pig, PWG miniature pig and Duroc were used. The ejaculated semen from the three pig species was diluted with same volume extender and added to LEY solution for freezing. The diluted semen was placed in 0.5 ml straws, and freezing was initiated by exposing the straws to liquid nitrogen (LN2) vapours for 10 min before placing them into LN2 for cryopreservation. A fter thawing, the sperm ability were assessed for viability (SYBR-14/PI staining), abnormality (Rose Bengal staining), and acrosome status (intactness, intensity and capacitation) (chlorotetracycline, CTC staining). The viability of frozen-thawed GalT KO pig sperm had no significant difference as compared with Duroc and PWG miniature pig sperm. However, The CTC pattern of frozen-thawed GalT KO cloned miniature pig spermatozoa showed significantly lower rates in F pattern and AR pattern (p<0.05) and significantly higher rates in B pattern than Duroc and PWG miniature pig (p<0.05). The abnormality of GalT KO cloned miniature pig sperm was significantly lower as compared to Duroc and PWG miniature pig sperm (p<0.05). In conclusion, GalT KO cloned miniature pig semen can be cryopreserved successfully and used for artificial insemination reasonably.

This study was conducted to examine the effect of oocyte donor age and micromanipulation medium on the development of mouse cloned embryos receiving cumulus cells. Mouse oocytes were obtained from 6 to 11 week-old mice BDF1 female mice(experiment 1) and cumulus cells were used as donor cells. Micromanipulation procedures for nuclear transfer(NT) were performed in FHM, M2 or Hepes-buffered TCM199(TCM199) medium(experiment 2). After nuclear transfer, the reconstructed oocytes were activated by 10 mM SrCl2 in Ca-free CZB medium in the presence of 5 μg/ml cytochalasin B for 5 h and cultured in KSOM medium for 4 days. In experiment 1, the survival rate of oocytes after injection of cumulus cells were significantly(p<0.05) lower in oocytes from 6～7 week-old mice(53.3%) than in oocytes from 8～9(80.9%) and 10～11 week-old mice(77.1%). In experiment 2, the survival rate of oocytes after cell injection were significantly(p<0.05) higher in FHM and M2 medium(71.7% and 76.9%) than in TCM199 medium (51.2%). The activation rates of cloned embryos were not different among the micromanipulation media. However, the embryos developed to blastocyst stage were significantly(p<0.05) higher in FHM medium(13.9%) than in M2 and TCM199 medium(0.0% and 0.0%). In conclusion, the present study suggest that oocytes from above 8 week-old mice are superior to oocytes from 6～7 week-old mice as a source of recipient cytoplasm and FHM is superior to M2 and TCM199 as a micromanipulation medium for mouse somatic cell cloning.

The overexpression of Phosphoprotein Enriched in Astrocytes (PEA15) gene is commonly found in human diabetic patients. The overexpression of this gene in skeletal muscle and fat tissues have been reported to cause insulin resistance, thereby impairing insulin stimulated glucose uptake. We introduced a gene of mouse PEA15 (mPEA15) and enhanced green fluorescent protein (EGFP) into fertilized one cell pig zygotes using microinjection, and produced a piglet that showed overexpression of mPEA15 in the muscle tissues and expression of EGFP in the ear tissues and hooves. RT-PCR RFLP, southern blot and FISH analysis showed that the tissues carried the transgene. Real-time RT-PCR and western blots demonstrated that PEA15 gene was overexpressed in the various tissues and muscle tissues, respectively. These facts suggest that expression vector system is normally expressed in the trnasgenic (TG) pigs. To use as animal diseases model for type 2 diabetes, further study is necessary to confirm whether diabetes occur in these TG pigs, especially insulin resistance.

This study was conducted to investigate the effects of donor cell treatments on the production of transgenic cloned piglets. Ear fibroblast cell obtained from NIH MHC Inbred minipig was used as control. The GalT knock-out/CD46 knock-in (GalT/CD46) transgenic cell lines were established and used as donor cells. The reconstructed GalT/CD46 embryos were surgically transferred into oviduct of naturally cycling surrogate sows (Landrace × Yorkshire) on the second day of standing estrus. Unlike control (1.2 kV/cm,, 75.4%), the fusion rate of the GalT/CD46 donor cells was significantly higher in 1.5 kV/cm, (84.5%) than that of 1.25 kV/cm, (20.2%) (p<0.01). When the number of the transferred embryos were more than 129, the pregnancy and delivery rates were increased to 13/20 (65%) and 5/20 (25%) compared to less than 100 group [1/6 (16.7%) and 0/6 (0%)], respectively. To analyze the effect of donor cell culture condition on pregnancy and delivery rates, the GalT/CD46 donor cells were cultured with DMEM or serum reduced medium. In serum reduced medium group, the pregnancy and delivery rates were improved to 8/12 (66.7%) and 5/12 (41.7%) compared to DMEM group [3/7 (42.9%) and 0/7 (0%)], respectively. In conclusion, it can be postulated that an appropriate fusion condition and culture system is essential factors to increase the efficiency of the production of transgenic cloned piglets.

Real-time PCR could help to provide answers to urgent questions about the incidence, prevalence, and epidemiology of currently emerging food-borne bacteria and diseases as identification and detection tools. The objective of this study was carried out to examine several critical parameters that must be optimized when converting from the ABI Prism 7000 SDS platform to the Cepheid SmartCycler Ⅱ so as to directly use the same primer and probe sequences. A lyophilized master mix-OmniMix HS bead, MgCl2 concentration, and PCR cycling conditions were evaluated so as to convert to a new platform, Smartcycler Ⅱ. The best optimal cycling conditions to detect Cronobacter sakazakii on SmartCycler Ⅱ were as follow: initial denaturation at 95℃ for 2 min followed by 45 cycles of 95℃ for 15 s, and 60℃ for 60 s using OmniMix HS bead contained 6 mM MgCl2 concentration. And the Ct value was 16.97 compared to 23.84 of Ct value in ABI Prism 7000 SDS. This result showed that when the several analytical parameters were taken the consideration for optimization, it could be performed assays between real-time PCR platforms. Also it is need of further study to develop the new single multiplex real-time PCR method for determining various Cronobacter spp. including three subspecies, too.

학습조직은 조직학습의 일련의 활동을 포함하고 이 활동이 원활히 이루어질 수 있도록 조직의 구조적인 측면을 통해 개인들이 지식을 공유하고 창조할 수 있는 능력을 함양할 수 있는 여건을 마련해 주는 조직을 의미하며, 학습조직은 미래 사회에서 조직이 갖춰야 할 또 다른 경쟁력인 혁신에도 기여할 수 있으리란 예상이 가능하게 됨에 따라 학습조직 구축에 대한 필요성이 대두되었다. 이에 학습조직을 성공적으로 운영하고 있는 기업의 운영사례를 제시하여 학습조직과 혁신이 조직의 장기적인 성과를 증진시킬 수 있도록 하기 위해서는 조직의 적응력 과 유연성을 강화시키는 것이 필요하다는 인식하에 본 연구는 연구자의 컨설팅 경험을 토대로 광주지역에서 중소기업의 학습조직을 성공적으로 운영하고 있는 기업을 선정하여 학습조 운영 및 학습활동, 학습조 성공사례를 제시하여 자발적인 학습활동을 높이고 학습 성과와 생산성 향상방안을 높이고자 하였다. 본 연구에서 보여준 운영사례에서 살펴볼 수 있듯이 조직구 성원들의 자발적인 학습조 활동을 통한 조직구성원들의 역량개발을 통해 생산성 향상과 기업의 가치제고를 위한 중소기업의 인적자원 개발이 아주 중요한 부분을 담당하고 있다고 보여진 바 중소기업의 적극적인 참여가 경영성과 향상에 크게 기여할 것으로 기대된다.