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GFP(Green Fluorescent Protein)가 발현되는 형질전환 닭의 생산
구본철,권모선,전익수,김태완 충남대학교 형질전환복제돼지연구센터 2004 논문집 Vol. No.8
본 연구에서는 VSV-G(vesicular stomatitis virus G glycoprotein)로 포장된 MoMLV(Moloney murine leukemia virus) retrovirus vector system을 이용하여 GFP가 발현되는 형질전환 닭을 생산하고자 하였다. GFP 유전자를 retroviral vector 내의 RSV(Rous sarcoma virus) Promoter의 조절 하에 도입한 후, GP293 세포주에서 virus 형태로 생산하였으며, 이 virus를 초원심분리로 고농축하여 stage X 계란의 배반엽 층에 주입하여 GFP가 발현되는 형질전환 닭을 생산하였다. 생산된 닭에서의 GFP의 발현은 epifluorescence stereomicroscope를 이용하여 확인하였다. 이 방법은 기존의 여러 형질전환 가금 방법에 비하여 기술적인 용이성과 경제성을 가지므로(Muramatsu, Park and Okumura 1998), 매우 효율적이고 주목할 만한 형질전환 가금 생산 방법으로 사료된다.
Adjuvant Glucocorticoids Therapy in Canine Mast Cell Tumor
Kim, Myung-jin,Lee, Jae-il,Kim, Young-suk,Son, Hwa-young,Jun, Moo-hyung,Park, Chang-sik,Kim, Myung-cheol 충남대학교 형질전환복제돼지연구센터 2007 논문집 Vol. No.10
구토와 식용부진을 3일전부터 나타내고,2년전부터 좌측 후지 상부에 종괴를 갖고 있는 12년령,8.0 kg,난소 제거 잡종 암캐 l두가 충남대학교 부속동물병원에 내원하였다. 환자는 fine-needle aspiration (FNA) 세포진단학 검사에 의하여 mast cell tumor로 진단하였다. Mast cell tumor 진단을 위한 WHO clinical staging system에 의하여 stage IIIa로 분류하였다 환자는 adjuvant corticosteroid 요법에 의하여 투약되었으며,축주의 요구에 의하여 완전 외과 절제는 실시하지 아니 하였다. 초기에 adjuvant corticosteroid 단독 요법에 의하여 종괴의 크기가 점차 감소하였으며,환자의 전신상태는 호전되었다. 그러나 그 후에는 더 이상 glucocorticoid에 반응을 하지 아니 하였으며,종괴의 크기가 증가하였고,2개월 후에 간헐적 구토와 심한 호흡곤란 때문에 안락사 하였다. 비장 종괴,십이지장 궤양,간 종괴와 후지 상부 근육부위에 침윤된 mast cell tumor가 부검 시에 발견되었다. Mast cell tumor는 grade I 또는 II에서 그리고 조직학적으로 well-differentiated된 상태에서 외과적 완전절제,방사선요법,adjuvant corticosteroid 요법,화학요법을 병용 하여야 양호한 예후를 나타낼 것으로 기대된다. A 12-year-old, 8.0 kg, spayed female, mixed-breed dog was presented to the Veterinary Medical Teaching Hospital of Chungnam National University (VMTH, CNU). That case has been growing up mass. in her left upper hindlimb about for 2 years and has showed vomiting and anorexia for 3 days. The patient was diagnosed with mast cell tumor on the basis of fine-needle aspiration (FNA) cytology techniques. According to World Health Organization clinical staging system for diagnosing mast cell tumors, it was classified into stage IIIa. The patient was treated by adjuvant corticosteroid therapy, but complete surgical excision was not achieved by owner's request. In the early stage of therapy, the size of the mass was gradually reduced with only adjuvant glucocorticoid therapy, so the patient's general condition was maintained well. But after 53 days later, the treatmant was not effective anymore and mass size was increased. Two months later, she was euthanized because of intermittent vomiting and severe respiratory distress. Splenic mass, duodenal ulceration, liver mass and infiltrated mast cell tumor in upper hindlimb muscle region were found in necropsy examination.
Yi, Y.J.,Park, C.S. 충남대학교 형질전환복제돼지연구센터 2004 논문집 Vol. No.8
This study wag carried out to investigate the vrious concentrations and exposure times of ethanol, one of many intracellular calcium elevating agents, and a sequential combination of ethanol(8%), cycloheximide (CHX,10 ㎍/ml), cytochalasin B (CCB, ㎍/ml) and 6-DMAP,2mM) to improve parthenogenetic activation and development of in vitro matured porcine oocytes. Cumulus-oocyte complexes(COCs) were matured in tissue culture medium (TCP) 199 for 44 h at 38.5℃, 5% CO₂ in air. Cumulus-free oocytes showing first polar body were activated by concentrations of 0, 5, 6, 7, 8, 9 and 10% ethanol for 10 min and exposure times of 0, 5, 8, 10, 12 and 15 min with 8% ethanol in HEPES buffered(25mM) NCSU-23 medium. Also, oocytes were activated with the NCSU-23 medium containing 8% ethanol for 10 min. After that, oocytes were incubated in the NCSU-23 medium supplemented with CHC, CCB, 6-DMAP, CCB +6-DMAP and CHX+CCB+6-DMAP for 3 h, respectively. Following activation, oocytes were transferred into the NCSU-23 medium containing 0.4% BSA for further culture of 20 and 144h at 38.5℃, 5% CO₂in air. The activation rates of oocytes were higher in 6, 7 and 8% ethanol concentration compared with 0, 5, 9 and 10% ethanol concentrations. Significantly, more oocytes (29.3-33.7) were activated in the exposure for 8, 10, 12 and 15 min than those in the exposure for 0 and 5 min, but there was no difference due to exposure to 8% ethanol for 8-15 min. Oocytes treated by chemical agents(40.5-70.5%) after exposure to ethanol significantly improved the rate of oocyte activation compared with ethanol alone(31.2%). The percentage of cleaved oocytes was higher in the ethanol + CHX +