用显微注射法制备携带Epstein-Barr病毒膜抗原基因--BLLF1的转基因小鼠

目的 :用显微注射法制备携带EB病毒 (Epstein Barrvirus,EBV)膜抗原 (membraneantigen ,MA)基因 BLLF1的转基因小鼠。方法 :对 38只昆明种小鼠进行超排卵 ,收集受精卵 ,将EBVMA基因 BLLF1显微注射到受精卵的原核内。将注射后成活且健康的受精卵移植到假孕母鼠的输卵管内使其发育直至分娩。PCR分析检测仔鼠基因组中转基因的整合。结果 :显微注射 6 77个受精卵 ,其中成活且健康的 32 0个 ,卵的成活率为 47.2 %。将其移植到 12只假孕母鼠的输卵管内 ,其中 2只鼠怀孕并产仔 12只 ,出生率为 3 .75 %。PCR分析 5只仔鼠基因组整合有BLLF1基因 ,整合率为 41.7%。结论 :携带EBVMA基因 BLLF1的转基因小鼠已制备成功

Production of the transgenic mice carrying Epstein-Barr virus membrane antigen gene-BLLF1 by microinjection

Objective: To produce the transgenic mice carrying Epstein Barr virus(EBV) membrane antigen (MA) gene BLLF1 by microinjection. Methods: 38 Kunming strain female mice were induced to give superovulation. The fertilized eggs were harvested and microinjected with EBV MA gene BLLF1 into their pronuclei. After microinjection, the survival and healthy fertilized eggs were implanted into the oviducts of pseudopregnant female mice to let them develop until delivery. PCR analysis was used to examine the integration of transgene in the offspring's genomes. Results: In this experiment, 677 fertilized eggs were microinjected and 320 of them survived and were healthy after microinjection.The survival rate was 47.2%. These eggs were implanted into the oviducts of 12 pseudopregnant female mice, of which two were pregnant and 12 offspring were born. The birth rate was 3.75%. PCR analysis showed that the genomes of 5 offspring were integrated with the BLLF1gene. The integration frequency was 41.7%. Conclusion: The transgenic mice carrying the EBV MA gene BLLF1 were successfully produced.