Vyskot B, Brzobohatý B, Karlovská L, Bezdĕk M
Institute of Biophysics, Czechoslovak Academy of Sciences, Brno.
Folia Biol (Praha). 1989;35(6):360-72.
The Drosophila genomic fragment Dm111 and the selectable dominant nptII gene were transferred via a Ti-vector into tobacco plants in order to check the structural and functional stability of transgenes in plants and their progeny. Southern blot analyses clearly showed that transgenes were integrated intact and did not suffer from any gross DNA rearrangements. Contrary to this structural stability, not all of the transgenic plants and their offspring displayed the original and stable expression of the nptII gene. The levels of the NPTII enzyme strongly varied in individual plants and did not depend on the copy number of the nptII gene. Both the non-stable nptII expression during the individual plant development in one original transgenic tobacco and some irregularities in segregation ratios after self-pollination indicated that epigenetic effects due to methylation of DNA modulated the expression of foreign genes in transgenic plants. This conclusion was supported by a spontaneous and 5-azacytidine-stimulated demethylation.
为了检测转基因在植物及其后代中的结构和功能稳定性,通过Ti载体将果蝇基因组片段Dm111和可选择的显性nptII基因导入烟草植株。Southern杂交分析清楚地表明转基因完整整合,未发生任何明显的DNA重排。与这种结构稳定性相反,并非所有转基因植株及其后代都表现出nptII基因的原始稳定表达。NPTII酶的水平在个体植株中差异很大,且不依赖于nptII基因的拷贝数。一株原始转基因烟草在个体植株发育过程中nptII表达不稳定,以及自花授粉后分离比例出现一些异常,这两者都表明DNA甲基化引起的表观遗传效应调节了转基因植物中外源基因的表达。这一结论得到了自发去甲基化和5-氮杂胞苷刺激去甲基化的支持。