Harris T J, Patel T, Marston F A, Little S, Emtage J S, Opdenakker G, Volckaert G, Rombauts W, Billiau A, De Somer P
Mol Biol Med. 1986 Jun;3(3):279-92.
cDNA clones of the mRNA coding for tissue-type plasminogen activator (t-PA) have been obtained and their nucleotide sequences compared to those reported previously. A gene coding for t-PA has been reconstructed and inserted into vectors for expression in prokaryotic cells. Relatively high levels of t-PA accumulated in inclusion bodies in Escherichia coli containing an optimized expression plasmid, but only a small proportion of the insoluble protein was recovered as active enzyme using a variety of solubilization procedures.
已获得编码组织型纤溶酶原激活剂(t-PA)的mRNA的cDNA克隆,并将其核苷酸序列与先前报道的序列进行了比较。已重建了编码t-PA的基因,并将其插入载体中以便在原核细胞中表达。在含有优化表达质粒的大肠杆菌中,相对高水平的t-PA积聚在包涵体中,但使用各种溶解方法,只有一小部分不溶性蛋白能作为活性酶回收。