CLONING OF THE CDNA-ENCODING HUMAN BRAIN TRYPSINOGEN AND CHARACTERIZATION OF ITS PRODUCT

被引：100

作者：

WIEGAND, U ^{[1
]}

CORBACH, S ^{[1
]}

MINN, A ^{[1
]}

KANG, J ^{[1
]}

MULLERHILL, B ^{[1
]}

机构：

[1] UNIV COLOGNE,INST GENET,D-50931 COLOGNE,GERMANY

来源：

GENE | 1993年 / 136卷 / 1-2期

关键词：

PCR AMPLIFICATION; DEGENERATED PRIMER DESIGN; MESSENGER-RNA EXPRESSION PATTERN; ALTERNATIVE SPLICING; IN VITRO TRANSLATION; NON-AUG START CODON; BETA-AMYLOID PRECURSOR PROTEIN;

D O I：

10.1016/0378-1119(93)90460-K

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

We designed degenerated oligodeoxyribonucleotide primers derived from amino acid (aa) sequences of the highly conserved active sites of mammalian serine proteases (SPs). These primers were used to selectively amplify, in polymerase chain reactions (PCRs), cDNA fragments coding for a SP. We used poly(A)(+)RNA from human brain to obtain cDNA fragments and amplified one cDNA encoding a novel SP. The full-length nucleotide (nt) sequence was identified by PCR and screening a genomic library in order to obtain the 5'-region. The deduced aa sequence shows a high degree of homology to trypsinogens, except for the first exon. In addition to this brain-specific trypsinogen, there exists a variant of the cDNA in pancreas, differing only in the nt sequence of the first exon. An active form of the trypsin was synthesized in vitro and purified by affinity chromatography using soybean trypsin inhibitor (STI) agarose to demonstrate the trypsin-specific interaction with a naturally occurring inhibitor of trypsins.

引用

页码：167 / 175

页数：9

共 38 条

[1] SUBCELLULAR FATE OF THE INT-2 ONCOPROTEIN IS DETERMINED BY CHOICE OF INITIATION CODON [J].