应用荧光原位杂交检测EB病毒BNLF-1基因在转基因小鼠子代染色体上的整合及其定位

应用荧光原位杂交技术研究了EB病毒潜伏膜蛋白基因(BNLF-1)在转基因小鼠子二代染色体上的整合及其定位。结果在两只子二代转基因小鼠中,分别观察80个和60个分裂相,出现杂交信号的核型分别为27和18个,检出率为33.8％和30％。转基因分别整合在14号染色体和10号染色体上。提示转基因BNLF-1已稳定整合到转基因小鼠的染色体上,并通过生殖细胞遗传给子代;推测转基因原代鼠的转基因整合可能是随机的多位点整合。

Mapping BNLF-1 Transgene on Transgenic Mouse Progeny Chromosomes by Fluorescence in situ Hybridization

In order to determine the integration and localization of BNLF-1 transgene of EB virus on transgenic mouse chromosomes, biotin-dUTP labelled DNA probes were localized to the progeny mouse chromosomes by using fluorescence in situ hybridization (FISH). Metaphase chromosomes were prepared from two second filial generation (F_2) mice. One showed clear hybridization signals on the long arm of chromosome 14 in 27 out of 80 metaphase (33.8%) examined, while for the other one, 18 were positive in 60 metaphases (30%) observed and the fluoresent spots were seen on long-ann band of chromosome 10. Results showed that BNLF-1 transgene had been stably integrated on the host chromosomes and transmitted to progeny by germ cells. This study suggested the deduction that the transgene is integrated at random, multiple sites in the founder transgenic mouse genome.