Chan W Y, Tease L A, Borjigin J, Chan P K, Rennert O M, Srinivasan B, Shupert W L, Cook R G
Laboratory of Protein Studies, Oklahoma Medical Research Foundation, Oklahoma City 73104.
Hum Reprod. 1988 Jul;3(5):677-85. doi: 10.1093/oxfordjournals.humrep.a136766.
Six human SP1 clones were isolated from a term placental cDNA library by immunological screening. All six cDNA clones cross-hybridized. However, at least two classes of cDNA could be distinguished, based on the presence or absence of an EcoRI site in the insert. Northern blot analysis of human term placental mRNA with all six cloned inserts demonstrated the presence of two mRNA species of 1.6 and 2.4 kb, respectively. The amino acid sequences of tryptic fragments of pure human SP1 protein were determined for confirmation of the identity of these cDNA clones. Using one of the cloned cDNA as probe, two and ten hybridizing clones were isolated from a human testicular cDNA library and a HeLa cell cDNA library, respectively. Southern blot analysis of these clones showed strong hybridization with the SP1 cDNA probe under high stringency, indicating the presence of highly homologous mRNA species in these tissues.
通过免疫筛选从足月胎盘cDNA文库中分离出六个人类SP1克隆。所有六个cDNA克隆都发生了交叉杂交。然而，根据插入片段中EcoRI位点的有无，至少可以区分出两类cDNA。用所有六个克隆插入片段对人足月胎盘mRNA进行Northern印迹分析，结果表明分别存在1.6 kb和2.4 kb的两种mRNA。测定了纯人SP1蛋白胰蛋白酶片段的氨基酸序列，以确认这些cDNA克隆的身份。用其中一个克隆的cDNA作为探针，分别从人睾丸cDNA文库和HeLa细胞cDNA文库中分离出两个和十个杂交克隆。对这些克隆进行Southern印迹分析，结果表明在高严谨度下它们与SP1 cDNA探针发生强杂交，表明这些组织中存在高度同源的mRNA种类。
