Molecular cloning and identification of a putative PKC epsilon cDNA from Limulus polyphemus brain
| Autor: | Maria L Leite Browning, Zheng Cao, Robert F. Newkirk, James G. Townsel, Easton A. Reid, Guatum Chaudhuri, Yilun Wang |
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| Rok vydání: | 2002 |
| Předmět: |
Central Nervous System
Male DNA Complementary Molecular Sequence Data Biology Molecular cloning Polymerase Chain Reaction General Biochemistry Genetics and Molecular Biology Sequence Analysis Protein Complementary DNA Horseshoe Crabs Animals Amino Acid Sequence General Pharmacology Toxicology and Pharmaceutics Cloning Molecular Peptide sequence Protein kinase C In Situ Hybridization Protein Kinase C Gene Library Base Sequence cDNA library Brain General Medicine biology.organism_classification Fusion protein Molecular biology Isoenzymes Molecular Weight Polyclonal antibodies Limulus biology.protein Female |
| Zdroj: | Life sciences. 72(8) |
| ISSN: | 0024-3205 |
| Popis: | The protein kinase C (PKC) family of enzymes is broadly distributed and has been implicated in a diverse array of cellular functions. Recent evidence supporting PKC involvement in the regulation of the Limulus choline cotransporter prompted us to clone PKC from a Limulus central nervous system (CNS) cDNA library. An Aplysia californica calcium independent PKC (Apl II) cDNA probe was used to screen the library and 5′ RACE SMART™ PCR was used to obtain the full-length sequence. The resulting cDNA, which included 5′ and 3′ nontranslation regions, was 4675 bp. Analysis of the encoded peptide sequence using the Swiss–prot database revealed at least 58% identity to PKCe. A commercial polyclonal antibody against PKCe was used in Western blots to positively label a 30 kDa protein from Limulus CNS and the expressed fusion protein of the encoded sequence. These data support the presence of a newly identified PKC-like homolog in Limulus which likely represents a PKCe equivalent. |
| Databáze: | OpenAIRE |
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