Shvartsman A L, Voronina O V, Gaĭtskhoki V S, Nadorshina O Kh, Timchenko L T
Mol Biol (Mosk). 1985 Mar-Apr;19(2):496-504.
Three clones carrying the sequences of ceruloplasmin gene were isolated from the library of EcoRI fragments of rat chromosomal DNA cloned in Charon 4A vector. These clones were identified using, i) plaque hybridization with [32P] cDNA transcribed from highly purified rat ceruloplasmin (Cp) mRNA; ii) blot hybridization of the restriction fragments of recombinant DNAs with Cp cDNA and iii) hybridization selection of Cp mRNA followed by its cell-free translation. Oligo (dT)-primed cDNA transcripts of Cp mRNA having different length as well as cloned Cp cDNA isolated from rat liver cDNA library were used as hybridization probes for the study of mRNA-coding segments of Cp gene. The length of inserts in recombinant DNAs varied from 7.5 up to 12.3 megadaltons. EcoRI-fragments of Cp gene were mapped within recombinant DNA and their homology to each other was studied.
从克隆于Charon 4A载体的大鼠染色体DNA的EcoRI片段文库中分离出三个携带铜蓝蛋白基因序列的克隆。这些克隆通过以下方法进行鉴定:i)用从高度纯化的大鼠铜蓝蛋白(Cp)mRNA转录的[32P] cDNA进行噬菌斑杂交;ii)重组DNA的限制性片段与Cp cDNA的印迹杂交;iii)Cp mRNA的杂交选择及其无细胞翻译。具有不同长度的Cp mRNA的寡聚(dT)引发的cDNA转录本以及从大鼠肝脏cDNA文库中分离的克隆的Cp cDNA用作杂交探针,用于研究Cp基因的mRNA编码区段。重组DNA中插入片段的长度从7.5到12.3兆道尔顿不等。Cp基因的EcoRI片段在重组DNA中进行定位,并研究它们之间的同源性。