Tang Z S, Wang X K, Li L C, Tian W Z
Institute of Genetics, Chinese Academy of Sciences, Beijing, China.
Yi Chuan Xue Bao. 2000;27(1):26-33.
Japonica rice cv 77170 was transformed with pBI222 carrying hygromycin phosphotransferase gene (hpt) by using biolistic bombardment and fertile transgenic rice plants were obtained. In T1 and most T2 generation the performance of hygromycin resistance was normal. Only in T2 progeny of SG-15 some lines showed decreased resistance. Mendelian inheritance of hygromycin resistance was showed as single dominant locus and proved by Southern blotting analysis in T1 and T2 generation of all transgenic plants. Multiple copies of hpt integrated into rice genome. These copies linked closely and inherited stablly. Msp I digested Southern blotting showed that methylation of hpt existed commonly in transgenic plants and their progeny, therefore uncomplete silencing of hpt in some T2 progeny of SG-15 was not associated with hpt methylation probably.
利用生物弹轰击法,将携带潮霉素磷酸转移酶基因(hpt)的pBI222载体导入粳稻品种cv 77170中,获得了可育的转基因水稻植株。在T1代和大多数T2代中,潮霉素抗性表现正常。仅在SG - 15的T2子代中,部分株系表现出抗性降低。潮霉素抗性呈孟德尔单显性位点遗传,并通过对所有转基因植株T1代和T2代的Southern杂交分析得以证实。hpt多拷贝整合到水稻基因组中,这些拷贝紧密连锁且稳定遗传。Msp I酶切Southern杂交显示,hpt甲基化在转基因植株及其子代中普遍存在,因此,SG - 15部分T2子代中hpt的不完全沉默可能与hpt甲基化无关。