Kang J, Wiegand U, Müller-Hill B
Institut für Genetik, Universität zu Köln, F.R.G.
Gene. 1992 Jan 15;110(2):181-7. doi: 10.1016/0378-1119(92)90646-7.
Serine proteases (SPs) are a family of physiologically important and versatile enzymes. We designed degenerated oligodeoxyribonucleotide primers derived from the consensus amino acid aa sequences of the active site of mammalian SPs, to selectively amplify in a polymerase chain reaction (PCR) cDNA fragments coding for SPs. We used poly(A)+ RNA from rat pancreas to obtain the cDNA. Two of the amplified cDNA fragments encode novel SPs. The full-length nucleotide sequence of both cDNAs was also obtained by PCR. The high degree of homology to trypsins and elastases suggests that the cDNAs encode a trypsin-like and an elastase-like SP, respectively. Both mRNAs were also found to occur, to a lesser extent, in spleen, as was the case for the mRNAs of other rat pancreatic SPs.
丝氨酸蛋白酶(SPs)是一类在生理上具有重要意义且功能多样的酶。我们根据哺乳动物丝氨酸蛋白酶活性位点的共有氨基酸序列设计了简并寡脱氧核糖核苷酸引物,以便在聚合酶链反应(PCR)中选择性扩增编码丝氨酸蛋白酶的cDNA片段。我们使用大鼠胰腺的聚腺苷酸加尾RNA(poly(A)+ RNA)来获取cDNA。其中两个扩增的cDNA片段编码新型丝氨酸蛋白酶。这两个cDNA的全长核苷酸序列也通过PCR获得。与胰蛋白酶和弹性蛋白酶的高度同源性表明,这两个cDNA分别编码一种类胰蛋白酶和一种类弹性蛋白酶的丝氨酸蛋白酶。正如其他大鼠胰腺丝氨酸蛋白酶的mRNA一样,这两种mRNA在脾脏中也有少量表达。