Jacob E
Nucleic Acids Res. 1980 Mar 25;8(6):1319-37. doi: 10.1093/nar/8.6.1319.
Double-stranded cDNA sequences were prepared from Xenopus laevis ovary poly A(+) RNA with AMV reverse transcriptase and nuclease S1. They were inserted into the plasmid pBR 322 after ligation with a Hind III linker and were cloned in E. coli strain X1776. Plasmid pools containing a cDNA insert were identified by Hind II restriction and hybridization of the DNA fragments with radiolabelled pBR 322 DNA. Hybridization of the positive pools with ovary RNA labelled in vitro led to the identification of cloned cDNA sequences which represent RNA species of high to intermediate abundance in the ovary. Positive clones were further challenged with in vitro labelled mitochondrial DNA and RNA from different developmental stages. One clone of mitochondrial origin has been detected. The hybridization characteristics of the cDNA sequences with the RNA probes from later developmental stages is discussed.
利用禽成髓细胞瘤病毒（AMV）逆转录酶和核酸酶S1从非洲爪蟾卵巢多聚腺苷酸（+）RNA制备双链cDNA序列。与Hind III接头连接后，将它们插入质粒pBR 322，并克隆到大肠杆菌菌株X1776中。通过Hind II酶切和用放射性标记的pBR 322 DNA对DNA片段进行杂交来鉴定含有cDNA插入片段的质粒文库。阳性文库与体外标记的卵巢RNA杂交，从而鉴定出代表卵巢中高至中等丰度RNA种类的克隆cDNA序列。用来自不同发育阶段的体外标记的线粒体DNA和RNA对阳性克隆进行进一步检测。检测到一个线粒体来源的克隆。讨论了cDNA序列与来自后期发育阶段的RNA探针的杂交特性。
