Edlund T, Ny T, Rånby M, Hedén L O, Palm G, Holmgren E, Josephson S
Proc Natl Acad Sci U S A. 1983 Jan;80(2):349-52. doi: 10.1073/pnas.80.2.349.
We have isolated a cDNA sequence coding for a part of human tissue plasminogen activator. mRNA coding for tissue plasminogen activator was partially purified, copied into double-stranded cDNA, and cloned into Escherichia coli. Two sets of partially overlapping oligodeoxynucleotide mixtures corresponding to all possible coding sequences for a known portion of the tissue plasminogen activator gene were prepared. One set was used as a probe to screen cDNA containing bacterial clones and both were used as probes in hybridization against purified plasmid DNA. Of 4,200 bacterial clones examined, 1 carried a plasmid that hybridized to both sets of oligonucleotides. This plasmid contained a 370-base-pair cDNA insert, which was shown by nucleotide sequence analysis to code for the cleavage site region in the one-chain form of the human tissue plasminogen activator.
我们已分离出一段编码人组织型纤溶酶原激活物部分序列的cDNA。编码组织型纤溶酶原激活物的mRNA被部分纯化，转录成双链cDNA，然后克隆到大肠杆菌中。制备了两组部分重叠的寡脱氧核苷酸混合物，它们对应于组织型纤溶酶原激活物基因已知部分的所有可能编码序列。一组用作探针筛选含细菌克隆的cDNA，两组都用作与纯化质粒DNA杂交的探针。在检测的4200个细菌克隆中，有1个携带的质粒与两组寡核苷酸都杂交。该质粒含有一个370个碱基对的cDNA插入片段，通过核苷酸序列分析表明其编码人组织型纤溶酶原激活物单链形式的裂解位点区域。
