Fluhr R, Fromm H, Edelman M
Gene. 1983 Nov;25(2-3):271-80. doi: 10.1016/0378-1119(83)90231-7.
All of the PstI restriction fragments of the chloroplast DNA of Nicotiana tabacum have been cloned in the plasmid vector pBR322. The cloned fragment sizes range from 0.8 to 26 kb, are stable, and can be amplified by chloramphenicol with varying efficiencies. Using these clones we have detailed a PstI physical map of the tobacco chloroplast genome. Selected clones of SalI, BamHI and PstI fragments were used to localize the map positions of the alpha, beta, and epsilon subunits of the chloroplast ATPase coupling factor, the large subunit of ribulosediphosphate carboxylase and the 32-kDal membrane protein. The gene products of these clones were characterized by RNA transcript sizing, immunoprecipitation of maxicell-directed protein synthesis, and hybrid-arrested translation.
烟草叶绿体DNA的所有PstI限制性片段已被克隆到质粒载体pBR322中。克隆片段大小在0.8至26 kb之间，是稳定的，并且可以用氯霉素以不同效率进行扩增。利用这些克隆，我们详细绘制了烟草叶绿体基因组的PstI物理图谱。选择的SalI、BamHI和PstI片段克隆用于定位叶绿体ATP酶偶联因子的α、β和ε亚基、核酮糖二磷酸羧化酶的大亚基以及32-kDal膜蛋白的图谱位置。这些克隆的基因产物通过RNA转录本大小测定、对最大细胞指导的蛋白质合成进行免疫沉淀以及杂交阻断翻译来进行表征。
