한우의 mt DNA cytochrome oxidase subunit I, II, 및 III complex지역의 유전적 다형현상을 제한효소를 이용하여 검출하였다. PCR primer 6종에 대하여 20가지 제한효소를 처리하였으며, Pst I, Pvu II, Rsa I, Eco RI, Bgl II, and Msp I 제한효소를 사용하여 유전적 변이를 검출하였다. 검출된 변이체와 한우의 성장과의 관련성을 조사한 결과 cytochrome oxidase subu

The purpose of this study was to investigate the development of bovine nuclear transfer (NT) embryos cultured in serum-free conditions. Bovine NT embryos cultured in various culture conditions were compared blastocyst development, total cell number and apoptosis using TUNEL assay. In experiment 1, blastocyst rates of NT embryos were significantly higher (P<0.01) in FBS (22.0％) and BSA (26.6％) groups than in PVA (6.3％) group. Total cell number was significantly higher in FBS (78.4±19.4) and BSA (90.9±29.1) groups than in PVA group (46.0±0.0). Apoptotic cell number was significantly fewer in FBS (3.1±1.4) and BSA (1.7±1.4) groups than in PVA group (7.0±20.0) However, all of results were not different between the FBS and BSA group. In experiment 2, blastocyst rates of NT embryos were significantly higher (P<0.05) in fatty acid free-BSA (FAF-BSA) group (26.8％) than in fraction V-BSA group (11.2％). Total cell number were somewhat higher in FAF-BSA group (89.8±30.7) than in fraction V-BSA group (88.1±19.3). Apoptotic cell number were somewhat fewer in FAF-BSA (1.7±1.5) group than in fraction V-BSA group (4.2±2.9). These findings suggest that serum free condition were effective for the in vitro development of bovine NT embryos. Therefore, we concluded that fatty acid free-BSA has beneficial effect in development bovine NT embryos and can be use as a serum substitute.

This study was performed to test the cellulose digestibility using the transgenic pigs harboring cellulose degradation gene D (CelD). After delivered offsprings between normal pig and transgenic swine, DNA was isolated from piglets tail for PCR analysis. In first generation, five out of 65 piglets showed CelD positive. Unfortunately, four CelD-positive pigs were died during growing, but one survived pig was used as a transgenic founder to produce F₁ descendents. Among 3 F₁ transgenic pigs produced, one died and the remaining two pigs were used to test the fiber digest efficiency. An assorted feed was composite of 5％ fiber with other ingredients. The feed of 3 kg per day was provided to the pigs including transgenic founders and littermate controls. The manure quantity was measured daily for a month, and all manures were dried for three days to analysis nitrogen, phosphate and fiber concentrations. The fiber digestion efficiencies of the transgenic F₁ pigs showed approximately 10％ higher than those of control pigs. Fiber digestion was not greatly improved in transgenic pigs as it had been expected approximately 30％. Nitrogen concentration of transgenic pig′s manure was slowly decreased compare to the control pigs. Because there were only two transgenic pigs tested, a large number of transgenic pigs may be necessary to obtain more reliable data. Breeding of animals to obtain sufficient transgenic pigs subjected for a further study is on progress. Taken together, this study demonstrated successful production of transgenic pigs with increase of cellulose digestibility in the porcine feed.

The present study were performed to analysis the hematocrit and the red blood cells content into the blood plasma of the transgenic pigs harboring recombinent human erythropoietin gene (rhEPO). Mouse whey acidic protein (mWAP) linked to rhEPO gene was microinjected into pronuclei of porcine one-cell zygotes. After delivered of offspring, PCR analyses identified one mWAP-rhEPO transgenic founder offspring(F/sub 0/). The first generation of transgenic pig (F/sub 0/) harboring mWAP-hEPO appeared to be a male, and the second generation (F₁) pigs were made by natural mating of F/sub 0/ with domestic swine, and male and female transgenic pigs (F₁) were identified by PCR. The blood samples from transgenic and normal pigs were collected for 50 days during lactation and were counted the red blood cell (RBC) numbers and Hematocrit (HCT) content into the blood. The transgenic pigs expressing rhEPO in their blood gave rise to higher RBC numbers and HCT contents than control animals. rhEPO was secreted both in the blood and milk of genetically engineered pigs harboring rhEPO gene. Therefore, this study provides a model regarding the production of transgenic pig carrying hEPO transgene for biomedical research.

The present study was conducted for the production of transgenic cloned cows by somatic cell nuclear transfer (SCNT) that secrete human prourokinase into milk. To establish an efficient production system for bovine transgenic SCNT embryos, the offset was examined of various conditions of donor cells including cell type, size, and passage number on the developmental competence of transgenic SCNT embryos. An expression plasmid far human prourokinase (pbeta-ProU) was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human prourokinase target gene into a pcDNA3 plasmid. Three types of bovine somatic cells including two adult cells (cumulus cells and ear fibroblasts) and fetal fibroblasts were prepared and transfected using a lipid-meidated method. In Experiment 1, developmental competence and rates of GFP expression in bovine transgenic SCNT embryos reconstructed with cumulus cells were significantly higher than those from fetal and ear fibroblasts. In Experiment 2, the effect of cellular senescence in early (2 to 4) and late (8 to 12) passages was investigated. No significant differences in the development of transgenic SCNT embryos were observed. In Experient 3, different sizes of GFP-expressing transfected cumulus cells [large (>30 ) or small cell (<30 )] were used for SCNT. A significant improvement in embryo development and GFP expression was observed when small cumulus cells were used for SCNT. Taken together, these results demonstrate that (1) adult somatic cells could serve as donor cells in transgenic SCNT embryo production and cumulus cells with small size at early passage were the optimal cell type, and (2) transgenic SCNT embryos derived from adult somatic cells have embryonic development potential.

We review observational evidence bearing on the formation of a prototypical large spiral galaxy, the Milky Way. New ground- and space-based studies of globular star clusters and dwarf spheroidal galaxies provide a wealth of information to constrain theories of galaxy formation. It appears likely that the Milky Way formed by an combination of rapid, dissipative collapse and mergers, but the relative contributions of these two mechanisms remain controversial. New evidence, however, indicates that initial star and star cluster formation occurred simultaneously over a volume that presently extends to twice the distance of the Magellanic Clouds.