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Publication : Molecular cloning, purification and in situ localization of human colon kallikrein.

First Author  Chen LM Year  1995
Journal  Biochem J Volume  307 ( Pt 2)
Pages  481-6 PubMed ID  7733886
Mgi Jnum  J:26651 Mgi Id  MGI:74084
Doi  10.1042/bj3070481 Citation  Chen LM, et al. (1995) Molecular cloning, purification and in situ localization of human colon kallikrein. Biochem J 307(Pt 2):481-6
abstractText  We have cloned and characterized a full-length cDNA encoding tissue kallikrein from a human colon carcinoma cell line (T84). The nucleic acid sequence of the colon kallikrein cDNA is identical to that of renal/pancreatic or tissue kallikrein cDNA. Reverse-transcription PCR followed by Southern-blot analysis using specific oligonucleotide probes showed expression of tissue kallikrein in human colon, pancreas and kidney. Tissue kallikrein mRNA was localized in glandular epithelial cells (goblet cells) in colon by in situ hybridization histochemistry. Human colon kallikrein was purified to apparent homogeneity by DEAE-Sepharose Cl-6B, aprotinin-affinity, and HQ/M perfusion chromatography. The purified colon kallikrein migrated as a broad, 40-45 kDa band in SDS/PAGE and was recognized by antibodies to human tissue kallikrein. The linear displacement curves for the colon kallikrein in an RIA were parallel with the human tissue kallikrein standard curve, indicating their immunological identity. The N-terminal sequence of the purified colon kallikrein matches completely with that of purified urinary or tissue kallikrein. These results indicate that human colon kallikrein is transcribed from the tissue kallikrein gene.
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