Molecular Cloning of the Oncofetal Isoform of the Human Pancreatic Bile Salt-dependent Lipase

摘要: Specific transcripts for bile salt-dependent lipase (BSDL), a 100-kDa glycoprotein secreted by the human pancreas, were immunodetected in BxPC-3 and SOJ-6 pancreatic tumoral cell lines. Sequencing of fragments, obtained mRNA reverse transcription amplification, confirmed presence BSDL these cancer cells. The protein was detected lysates cells, where it mainly associated with membranes. Only minute amount enzyme culture media. Immunofluorescence studies demonstrated that colocates p58 Golgi suggested may be sequestrated within compartment. These results is expressed cells cannot (or least very poorly). Subsequently, cDNA covering entire sequence transcription-polymerase chain reaction. this indicated N-terminal domain encoded exons 1–10 identical to normal pancreas. However, corresponding exon 11, which should code 16 tandem-repeated mucin-like sequences BSDL, deleted 330 base pairs (bp) only 6 repeated sequences. We conclude truncated variant would its oncofetal form, referred as feto-acinar protein. then investigated whether deletion bp affected secretion For purpose, mature form cloned into an expression/secretion vector transfected CHO-K1 Results isolated from level significantly higher than observed Consequently, retention might not result inherent properties