외부에서 투여된 열자극, 알콜 및 생리적 염과 같은 환경 스트레스는 체내 각 부위에서 스트레스단백질(열자극단백질, HSP)을 생성하게 된다. 본 연구에서는 비소가 흰쥐 대동맥의 수축에 미치는 영향을 조사하기 위해 스트레스단백질의 발현과 대동맥의 수축력의 변화와 이들과의 관계를 알아보고자 실험을 실시하였다 적출한 혈관은 organ bath에 담가 0, 0.5, 1, 2,및 4 mM As를 처리한 후 1, 3, 및 8시간 뒤에 KCI(55 mM)에 대한 수

Further development of reconstructed embryos may be dependent upon the synchronization of donor nucleus and recipient cytoplasm at cell fusion, To control the synchronization of donor and recipient cells, the enucleated MII arrested oocytes are artificially stimulated prior to embryo reconstruction. Destruction of cyclin B results in the exit of cells from M-phase of cell cycle. This study was designed to investigate the effects of single or combined stimulation affected cyclin B1 mRNA and protein levels in mouse oocytes. The oocyte activation was induced by 7% ethanol or 10/ Ca-ionophore without (single) or with (combined) 10/ cycloheximide. Competitive quantitative PCR for cyclin Bl mRNA and western blot analysis for cyclin B1 protein was preformed in mouse oocytes. Cyclin B1 mRNA level was significantly reduced in single (P<0.05) and combined (P<0.05) stimulation groups. However, this level did not change in non-activated group and increased in intact group. Cyclin B1 protein level was also significantly reduced in both single (P<0.05) and combined (P<0.05) stimulation groups. In conclusion, single and combined stimulation induces the degradation of cyclin B1 mRNA and protein after activation in enucleated mouse oocytes.

The role of heat shock proteins in shielding organism from environmental stress is illustrated by the large-scale synthesis of these protein by the organism studied to date. However, recent evidence also suggests an important role for heat shock protein in fertilization and early development of mammalian embryos. Effects of elevated in vitro temperature on in vitro produced bovine embryos were analysed in order to determine its impact on the expression of heat shock protein 70 (HSP70) by control and frozen-thawed after in vitro fertilization (IVF) or nuclear transfer (NT). The objective of this study was to assess the developmental potential in vitro produced embryos with using of the various containers and examined expression and localization of heat shock protein 70 after it's frozen -thawed. For the vitrification, in vitro produced embryos at 2 cell, 8 cell and blastocysts stage after IVF and NT were exposed the ethylene glycol 5.5 M freezing solution (EG 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min, and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid, cryo-loop. However, survival rates by straw were relatively lower than other containers. Only, nuclear transferred embryos survived by using cryo-loop. After IVF or NT, in vitro matured bovine embryos 2 cell, 8 cell and blastocysts subjected to control and thawed conditions were analysed by semiquantitive reverse transcription polymerase chain reaction methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNA were higher thawed embryos than control embryos. Immunocytochemistry used to localization the hsp70 protein in embryos. Two, 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some embryos exposed frozen-thawed. However, under control condition, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform in distribution.

Autographa californica 핵다각체병 바이러스(AcNPV)의 다각체 단백질과 초록색 형광 단백질의 융합단백질의 특성을 분석하였다. 초록색 형광 단백질 유전자는 AcNPV의 완전한 다각체 단백질 유전자의 앞쪽과 뒤쪽에 융합하여 다각체 단백질 유전자의 프로모터 조절하에 도입하였다. 이렇게 작성된 재조합 바이러스를 각각 Ac-GFPPOL 또는 Ac-POLGFP이라고 명명하였다. 이들 재조합 바이러스에 의해 감염된 곤충세포주에서는 56kDa의 융합단백질이 발현되었다. 한편, 흥미롭게도 재조합 바이러스 Ac-POLGFP에 의해 감염된 세포주에서는 초록색 형광이 핵내에서만 다각체 유사 granular particle 형태로 관찰되었다. 반면에 Ac-GFPPOP에 의해 감염된 세포도주에서는 대부분 핵내에 존재하였지만, 세포질과 핵 모두에서 초록색 형광을 관찰할 수 있었다. 그러나 발현된 융합단백질은 분명히 다각체단백질을 포함하고 있음에도 다각체는 형성하지 않았다. 이러한 결과들은 융합단백질에서 다각체단백질의 위치와 관련이 있는 것으로 보여진다.

The efficiency of transgenic livestock animal production may be improved by early selection of transgenci preimplantation embryos. To examine the possibility of GFP gene as a non-invasive marker for the early screening of transgenic embryo, the GFP gene was microinjected into rabbit zygotes and the later stages of preimplantation embryos were examined for the expression of GFP. The presence of injected DNA was detected by PCR analysis and the expression of GFP was detected by observing green fluorescence in embryos under a fluorescent microscope. Out of 108 GFP gene-injected rabbit zygotes, seventy three(67.6%) were fluorescence-positive. When 11 fluroresecence-positive blastocysts were analyzed for the presence of GFP gene by PCR, 6(54.5%) were positive, and all of the 8 flrouescence-negative blastocysts were also negative by PCR. The results indicate that the screening of transgene in rabbit embryos by PCR analysis and GFP detection could be a promising method for the preselection of transgenic embryos.

남부지방에서 분리한 곤충병원성곰팡이 Beauveria bassiana GY1-17균주를 이용하여 산림해충인 오리나무잎벌레(Agelastica coerulea), 밤나무혹나방(Meganola melancholica), 회양목명나방(Glyphodes perspectalis), 잔디해충인 등얼룩풍뎅이(Blitopertha orientalis), 채소해충인 배추좀나방(Plutella xylostella) 및 거세미나방(Agrotis segetum)의 생물적 방제 가능성을 알아보기 위하여 실험한 결과, 오리나무잎벌레와 배추좀나방유충은 7.0~2.0 conidia/ml 농도에서 처리 7일과 5일후 100%의 치사율을 나타내었다. 밤나무혹나방유충은 0.03875~3.1 conidia/ml 처리에서 66.7~100%의 높은 치사율을 보였으나 회양목명나방유충은 2.0~2.0conidia/ml 처리에서도 전혀 치사되지 않았다. 등얼룩풍뎅이유충은 3.7 conidia/ml 농도에서 46.7%의 치사율을 보였고, 거세미나방유충은 2.5 conidia/ml 농도에서 63.3% 치사율을 나타내었다.