Knöchel Walter, Bladauski Dieter
Institut für Molekularbiologie und Biochemie, Freie Universität Berlin, Arnimallee 22, D-1000, Berlin 33.
Wilehm Roux Arch Dev Biol. 1981 Mar;190(2):97-102. doi: 10.1007/BF00848402.
Nuclear poly(A) RNA was isolated from gastrula and early tadpole stages ofXenopus laevis, transcribed into cDNA and integrated as double stranded cDNA by the G-C joining method into the Pst cleavage site of plasmid pBR 322. After cloning inE. coli strain HB 101 the clone libraries were hybridized toP labelled cDNA derived from nuclear poly(A) RNA of the two different developmental stages. About 20% of the clones gave a positive hybridization signal thus representing RNA molecules of high and medium abundance. From these clones, some individual clones were identified containing sequences which are not present at the oocyte and gastrula stages but which are transcribed at the early tadpole stage of embryonic development.
从非洲爪蟾的原肠胚和早期蝌蚪阶段分离出核聚腺苷酸RNA,转录成cDNA,并通过G-C连接法作为双链cDNA整合到质粒pBR 322的Pst切割位点。在大肠杆菌HB 101菌株中克隆后,将克隆文库与源自两个不同发育阶段的核聚腺苷酸RNA的P标记cDNA杂交。约20%的克隆给出阳性杂交信号,因此代表高丰度和中等丰度的RNA分子。从这些克隆中,鉴定出一些单个克隆,其包含在卵母细胞和原肠胚阶段不存在但在胚胎发育的早期蝌蚪阶段转录的序列。