A previously identified rat cDNA (S1) that shares 78% nucleotide homology and a predicted 92% amino acid sequence homology with human EF-1$ alpha$ was used to express S1 in E. coli and to generate a polyclonal antibody to pS1. A recombinant plasmid pGEX-2T-S1 was constructed, containing the glutathione S-transferase gene. The expressed fusion protein was purified and digested with thrombin to produce a recombinant S1 protein (rpS1) containing slightly modified N-terminus. Purified rpS1 was used to raise a rabbit antiserum which recognized rpS1 on immunoblots. A polyclonal antiserum to EF-1$ alpha$ failed to react with rpS1. Similarly the anti-rpS1 does not recognize EF-1$ alpha$ on immunoblots. Anti-rpS1 therefore is able to distinguish pS1 from EF-1$ alpha$ despite their extensive amino acid sequence homology. Anti-rpS1 and anti-Ef-1$ alpha$ will be used to study the similarities and differences between pS1 and EF-1$ alpha$ in vivo and in vitro.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.61334 |
| Date | January 1992 |
| Creators | Liu, Catherine Heung Luen |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Master of Science (Department of Anatomy.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001307452, proquestno: AAIMM80353, Theses scanned by UMI/ProQuest. |
Page generated in 0.002 seconds