Bock S C, Wion K L, Vehar G A, Lawn R M
Nucleic Acids Res. 1982 Dec 20;10(24):8113-25. doi: 10.1093/nar/10.24.8113.
A partial cDNA clone for human antithrombin III (ATIII) was obtained by screening a cDNA library prepared from size fractionated liver RNA with a pool of eight 16-base long synthetic DNA fragments whose sequence was determined from protein sequence data. A fragment of the partial cDNA clone was used to enrich RNA for ATIII messages, and cDNA clones encoding the entire ATIII structural gene were identified. The complete nucleotide and predicted amino acid sequences of human ATIII and its 32 residue signal peptide are reported, and provide further opportunity to compare the ATIII primary structure with corresponding regions from homologous proteins, alpha 1-antitrypsin and ovalbumin. Plasmids in which the structural genes for mature and pre-ATIII were linked to the E. coli trp promoter-operator support the synthesis of human antithrombin III and pre-antithrombin III in bacteria.
通过用一组八个16碱基长的合成DNA片段筛选从肝脏分级分离的RNA构建的cDNA文库，获得了人抗凝血酶III（ATIII）的部分cDNA克隆，这些合成DNA片段的序列是根据蛋白质序列数据确定的。部分cDNA克隆的一个片段用于富集ATIII信使RNA，并且鉴定了编码整个ATIII结构基因的cDNA克隆。报道了人ATIII及其32个残基信号肽的完整核苷酸和预测氨基酸序列，这为将ATIII一级结构与同源蛋白α1-抗胰蛋白酶和卵清蛋白的相应区域进行比较提供了更多机会。其中成熟和前ATIII结构基因与大肠杆菌色氨酸启动子-操纵子相连的质粒支持在细菌中合成人抗凝血酶III和前抗凝血酶III。
