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RNA 10, 277–286. Localization of a promoter in the putative internal ribosome entry site of the Saccharomyces cerevisiae TIF4631 gene. 2004

Verge´, V., VonLanthenm, M., Masson, J.M., Trachsel, H., and Altmann, M.

Notes: Researchers cloned the Photinus and Renilla luciferase ORFs into the pSP64 Poly(A) Vector to create a dual-reporter vector named SP6P. A similar vector, SP6R.4G(-508/-3).P, was created in which a 5´ untranslated region from the Saccharomyces cerevisiae TIF4631 gene was cloned between the two reporter genes. These two vectors were used to transform yeast strains. The resultant transformants were lysed using Passive Lysis Buffer and a modified lysis procedure.   Lysates were analyzed for luciferase activities using the Dual-Luciferase® Reporter Assay System and a TD20/20 luminometer. The researchers also cloned and sequenced the 5´  untranslated region of TIF4631 by using a RACE-PCR technique followed by cloning the PCR amplimers into the pGEM®-T Vector. (2845)

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J. Biol. Chem. 274, 23910-23915. Deletions of specific substitutions of a few residues in the NH2-terminal region (Ala3 to Thr9) of sarcoplasmic reticulum Ca2+-ATPase cause inactivation and rapid degradation of the enzyme expressed in COS-1 cells. 1999

Daiho, T., Yamasaki, K., Suzuki, H., Saino, T., Kanazawa, T.

Notes: Wildtype and mutant Ca2+-ATPase cDNAs were cloned into the pSP64 Poly(A) Vector and expressed in the TNT® Reticulocyte Lysate System in the presence of the Canine Pancreatic Microsomal Membranes. The membranes were spun out of the reaction and analyzed by SDS-PAGE. Both wildtype and mutant proteins localized to the membranes. (1250)

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J. Neurosci. 17, 181-189. Conservation of topology, but not conformation, of the proteolipid proteins of the myelin sheath. 1997

Gow A., Gragerov A., Gard A., Colman D.R., Lazzarini R.A.

Notes: All cDNAs to be translated in vitro were subcloned into the pSP64 Poly(A) Vector. The vector was linearized and RNA transcribed from the SP6 promoter with the Riboprobe® in vitro Transcription System. The in vitro transcribed RNA was translated in the Rabbit Reticulocyte Lysate in the presence or absence of Canine Pancreatic Microsomal Membranes (CMMs). N-linked glycosylation sequences were engineered into the proteolipid protein to determine the orientation of the domains of the protein in the membrane. (1114)

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