Open AccessMolecular cloning and nucleotide sequence of rat lingual lipase cDNA
Author(s) -
Andrew Docherty,
Mark Bodmer,
Sarojani Angal,
Robert Verger,
C. Rivière,
Peter A. Lowe,
A. Bruce Lyons,
J.S. Emtage,
Timothy Harris
Publication year - 1985
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/13.6.1891
Subject(s) - biology , complementary dna , nucleic acid sequence , lipase , microbiology and biotechnology , peptide sequence , biochemistry , molecular cloning , amino acid , cdna library , polyclonal antibodies , triacylglycerol lipase , enzyme , antibody , genetics , gene
Purified rat lingual lipase (EC3113), a glycoprotein of approximate molecular weight 52,000, was used to generate polyclonal antibodies which were able to recognise the denatured and deglycosylated enzyme. These immunoglobulins were used to screen a cDNA library prepared from mRNA isolated from the serous glands of rat tongue cloned in E. coli expression vectors. An almost full length cDNA clone was isolated and the nucleotide and predicted amino acid sequence obtained. Comparison with the N-terminal amino acid sequence of the purified enzyme confirmed the identity of the cDNA and indicated that there was a hydrophobic signal sequence of 18 residues. The amino acid sequence of mature rat lingual lipase consists of 377 residues and shares little homology with porcine pancreatic lipase apart from a short region containing a serine residue at an analogous position to the ser 152 of the porcine enzyme.