Potrykus I, Paszkowski J, Saul M W, Petruska J, Shillito R D
Mol Gen Genet. 1985;199(2):169-77. doi: 10.1007/BF00330255.
Two clones of N. tabacum, transformed to kanamycin resistance by direct transfer to protoplasts of a hybrid gene, consisting of the protein coding region from the bacterial gene for aminoglycoside phosphotransferase under the control of 5'/3' expression signals from cauliflower mosaic virus gene VI, in the bacterial plasmid pUC8, have been subjected to a detailed genetic crossing analysis accompanied by Southern blot analysis and enzyme activity assays of representative offspring. The genetic data obtained from large populations of R1/F1 and R2/F2 offspring as well as from more than 20 subclones of each of the original of the hybrid gene was stably integrated into chromosomal DNA of the original transformants, (b) that the gene normally was stably maintained during clonal proliferation, (c) that normally it is transmitted in a regular fashion (with exceptions) to sexual offspring, and (d) that it is inherited as a single dominant trait. Data from DNA hybridisation and enzyme assays confirm this interpretation. The functional gene is integrated together with several non-functional copies and bacterial plasmid sequences, which are inherited as one block together with the functional gene.
两个烟草克隆体通过将由花椰菜花叶病毒基因VI的5'/3'表达信号控制的、来自细菌氨基糖苷磷酸转移酶基因的蛋白质编码区与细菌质粒pUC8中的杂合基因直接导入原生质体而转化为对卡那霉素具有抗性。对这两个克隆体进行了详细的遗传杂交分析,并对代表性后代进行了Southern印迹分析和酶活性测定。从大量R1/F1和R2/F2后代以及每个原始杂交基因的20多个亚克隆中获得的遗传数据表明:(a) 杂合基因稳定地整合到原始转化体的染色体DNA中;(b) 该基因在克隆增殖过程中通常能稳定维持;(c) 该基因通常以规则的方式(有例外情况)传递给有性后代;(d) 它作为单一显性性状遗传。DNA杂交和酶分析的数据证实了这一解释。功能基因与几个无功能的拷贝以及细菌质粒序列一起整合,它们与功能基因作为一个整体遗传。