title: Cloning of the cDNA for rabbit L-selectin and expression of recombinant protein with a kinase target site to facilitate radiolabeling.
abstract: The cDNA encoding rabbit L-Selectin has been cloned from a cDNA library, utilizing a PCR-derived probe. It encodes a peptide of 377 amino acids, including a signal peptide of 38 amino acids. Sequence analysis demonstrated extensive homology with L-Selectin's from other species. Recombinant rabbit L-Selectin protein was expressed in eukaryotic cells in a chimeric construct incorporating the entire extracellular portion of the protein coding region, a phosphokinase target site to allow high activity radiolabeling with 32P, and the constant region of the heavy chain of human IgG1 to facilitate purification and detection. Amino-terminal peptide sequencing of recombinant L-Selectin confirmed that the signal peptide had been removed at the expected site.
authors: Department of Internal Medicine, University of Michigan, Ann Arbor 48109-0531, USA.
authors: J
authors: J
authors: Qian
authors: X
authors: X
authors: Huang
authors: R M
authors: RM
authors: Marks
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006801:Humans; D019041:L-Selectin / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010761:Phosphorus Radioisotopes; D010770:Phosphotransferases / Q000378:metabolism; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D010761:Phosphorus Radioisotopes; D011994:Recombinant Proteins; D019041:L-Selectin; D010770:Phosphotransferases