Elsevier

Gene

Volume 136, Issues 1–2, 22 December 1993, Pages 167-175
Gene

Cloning of the cDNA encoding human brain trypsinogen and characterization of its product

https://doi.org/10.1016/0378-1119(93)90460-KGet rights and content

Abstract

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 demon- strate the trypsin-specific interaction with a naturally occurring inhibitor of trypsins.

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  • Cited by (0)

    Present addresses: Fachschule für Technik, Heegestr. 14, D-45897 Gelsenkirchen, Germany. Tel. (49-209) 59-919.

    ∗∗

    Present addresses: Diagen GmbH M.-Volmer-Str. 4, D-40724 Hilden, Germany. Tel. (49-2103) 892-263.

    3

    e-mail; [email protected]

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