节点文献
家兔个体表达系统的建立
Production of Transgenic Rabbits by Micro Injection
【摘要】 将乙肝表面抗原基因及人生长激素基因用微注射法转入家兔受精卵发育成为有整合、有表达的当代兔,并繁殖出了有外源基因存在、有表达的子代兔。转基因所用的质粒是含(1)乙肝病毒表面抗原S基因与小鼠MT启动子的pMTSA。(2)乙肝病毒核心抗原基因和乙肝病毒S基因启动子的pHBV(?)和(3)含有人生长激素基因SV40启动子和小鼠MT启动子的pSMGH。质粒均切成线性分子后再注射。从757个微注射并移植的受精卵经发育娩出得101只仔兔。其中有57%具有整合的微注射基因。用ELISA法测出28只转基因兔内有8只的血清中有表达的乙肝病毒表面抗原,占总数约30%。转基因兔与转基因兔和其与正常兔交配获得的子代中有83%含有转基因,有15%在血清中可测出乙型肝炎表面抗原。对转入人生长激素基因的兔也作了整合和表达的检测。
【Abstract】 The feasibility of using whole animal instead of bioreactor in genetic engineering has been investigated with transgenic domestic rabbits. The gene chosen is the surface gene (S gene) of hepatitis B virus. Two plasmids were specifically constructed for this purpose, pHBV3.0 contains the promoter pre 5 gene and a part of c gene of the virus; while MT-SA, the S gene and mouse MT promotor. These plasmids were made linear by suitable restiction endonuclease before they were transferred into maleprounclei by means of microinjection. From 757 mi-croinjected and transplanted fertilized eggs 101 rabbits were obtained. 57% of these animals were found with intergrated microinjected genes. 28 of the transgenic animals were tested for the presence of the surface antigen of the virus in the serum by ELISA method. 8 animals were found positive, approximately 30% of the tested transgenic animals. The second generation transgenics were obtained either by first generation ransgenics crossed with non-trans-genics or transgenics. Among them 73% contained the transgene and 15% had the surface antigen in the serum.Some expreiments were also carried out with human growth hormone gene.
【Key words】 Transgenic rabbit; pHBV3.0 pMT-SA; pSMGH; Microinjection; Integration; Expression;
- 【文献出处】 遗传学报 ,Acta Genetica Sinica , 编辑部邮箱 ,1993年02期
- 【被引频次】13
- 【下载频次】45