Landau N R, St John T P, Weissman I L, Wolf S C, Silverstone A E, Baltimore D
Proc Natl Acad Sci U S A. 1984 Sep;81(18):5836-40. doi: 10.1073/pnas.81.18.5836.
A cDNA library was prepared from a terminal deoxynucleotidyltransferase-containing thymoma in the lambda phage vector lambda gt11. By screening plaques with anti-terminal transferase antibody, positive clones were identified of which some had beta-galactosidase-cDNA fusion proteins identifiable after electrophoretic fractionation by immunoblotting with anti-terminal transferase antibody. The predominant class of cross-hybridizing clones was determined to represent cDNA for terminal transferase by showing that one representative clone hybridized to a 2200-nucleotide mRNA in close-matched enzyme-positive but not to enzyme-negative cells and that the cDNA selected a mRNA that translated to give a protein of the size and antigenic characteristics of terminal transferase. Only a small amount of genomic DNA hybridized to the longest available clone, indicating that the sequence is virtually unique in the mouse genome.
从含末端脱氧核苷酸转移酶的胸腺瘤中制备了λ噬菌体载体λgt11的cDNA文库。通过用抗末端转移酶抗体筛选噬菌斑，鉴定出阳性克隆，其中一些克隆的β-半乳糖苷酶-cDNA融合蛋白在经抗末端转移酶抗体免疫印迹进行电泳分离后可被识别。通过显示一个代表性克隆与酶阳性但非酶阴性细胞中2200个核苷酸的mRNA杂交，且该cDNA选择了一种可翻译出具有末端转移酶大小和抗原特性的蛋白质的mRNA，确定主要的交叉杂交克隆类群代表末端转移酶的cDNA。只有少量基因组DNA与最长的可用克隆杂交，表明该序列在小鼠基因组中实际上是独特的。
