Molecular cloning and expression of cDNA for rat pancreatic cholesterol esterase.

摘要: A full-length cDNA complementary to the rat pancreatic cholesterol esterase mRNA was isolated by screening a expression library in λgt11 vector with antibodies against porcine esterase. The is 2050 bp length and contains an open reading frame coding for protein of 612 amino acids. 20-amino acid hydrophobic leader sequence predicted, based on position first ATG initiation codon upstream from sequenced terminus subcloned into mammalian vector, pSVL, transfection studies. Expression COS cells resulted production bile salt-stimulated Comparison other proteins revealed that identical lysophospholipase. primary structure displayed no significant homology lipases, although putative lipid interfacial recognition site G-X-S-X-G present sequence. However, 63-amino-acid domain which highly homologous active serine esterases. These data suggest may be member supergene family. Analysis also repetitive enriched Pro, Asp, Glu, Ser, Thr residues at C-terminal end protein. This reminiscent PEST-rich sequences short-lived proteins, suggesting have short half-life vivo. Northern blot hybridization showed liver this synthesized cells.