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Citations Search

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Genes Dev. 12(15), 2293-2304. Switch from translation to RNA replication in a positive-stranded RNA virus. 1998

Gamarnik, A.V. and Andino, R.

Notes: Authors used the Rabbit Reticulocyte Lysate System to investigate the interaction between transcription and translation directed by poliovirus. Thirty-five microliters of lysate were supplemented with 4µg of a poliovirus-infected S10 HeLa extract containing the partially purified viral polymerase, and translation was monitored using a poliovirus replicon driving luciferase expression. RNA synthesis was measured by incorporation of 32P-UMP into the poliovirus RNA. When ribosomes were actively translating, RNA synthesis was undetectable, but when cycloheximide was added to the reactions, the viral polymerase was able to incorporate the labeled nucleotide. (2172)

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J. Biol. Chem. 273, 35362-35370. The 72-kDa component of signal recognition particle is cleaved during apoptosis. 1998

Utz, P. J., Hottelet, M., Le, T. M., Kim, S. J., Geiger, M. E., van Venrooij, W. J., Anderson, P.

Notes: The authors incubated Canine Pancreatic Microsomal Membranes (CMMs) with various caspases to show cleavage of SRP 72. The treated membranes were incubated with β-lactamase made in Rabbit Reticulocyte Lysate and the effect of cleavage of SRP 72 on processing/transport of β-lactamase was evaluated. (0210)

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EMBO J. 16, 659-671. A small heat shock protein stably binds heat-denatured model substrates and can maintain a substrate in a folding-competent state. 1997

Lee, G. J. , Roseman, A. M. , Saibil, H. R. , Vierling, E.

Notes: The Quantilum™ Recombinant Luciferase was heat denatured in the presence of heat shock protein hsp 18.1 and the renaturation of the protein was assayed in with the Luciferase Assay System. The denaturation was performed in either Rabbit Reticulocyte Lysate or Wheat Germ Extract in the presence or absence of added ATP. (0809)

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J. Biol. Chem. 272, 16934-16939. Activating transcription factor 2 (ATF2) down-regulates hepatitis B virus X promoter activity by the competition for the activating protein 1 binding site and the formation of the ATF2-Jun heterodimer 1997

Choi, C. Y. , Choi, B. H. , Park, G. T. , Rho, H. M.

Notes: The AP-1 and CREB Consensus Oligonucleotides were used for gel shift assays with transfected HepG2 cell extracts, E.coli extracts containing eukaryotic transcription factors and a truncated c-jun protein expressed with Rabbit Reticulocyte Lysate. (1343)

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J. Biol. Chem. 272, 255-261. Arginine-specific regulation mediated by the Neurospora crassa arg-2 upstream open reading frame in a homologous, cell-free in vitro translation system. 1997

Wang, Z. , Sachs, M. S.

Notes: Luciferase reporter constructs containing either the luc or luc+ gene (from pSP-luc+NF Fusion Vector) were used in in vitro transcription to produce capped mRNA. Luciferase was produced by in vitro translation using either neurospora extract or Promega's Rabbit Reticulocyte Lysate, Nuclease Treated and detected using the Luciferase Assay Reagent. (0208)

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Blood 90, 372-381. Calreticulin biosynthesis and processing in human myeloid cells: Demonstration of signal peptide cleavage and N-glycosylation. 1997

Denning, G.M., Leidal, K.G., Holst, V.A., Iyer, S.S., Pearson, D.W., Clark, J.R., Nauseef, W.M. and Clark, R.A.

Notes: PolyA+ RNA from HL-60 cells was in vitro translated using the Rabbit Reticulocyte Lysate System in the presence or absence of Canine Microsomal Membranes, then immunoprecipitated and analyzed by SDS-PAGE and fluorography. The calreticulin protein was presumably glycosylated by the membranes. The same results were obtained with in vitro transcripts of the calreticulin. (1617)

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EMBO J. 16, 2703-2716. Cell-free synthesis and assembly of connexins into functional gap junction membrane channels. 1997

Falk, M.M., Buehler, L.K., Kumar, N.M. and Gilula, N.B.

Notes: Several different gap junction channel subunit isotypes, also known as connexins, were synthesized in vitro in the presence of Canine Microsomal Membranes. Translation reactions were spun through a sucrose cushion to separate the membrane fraction from the supernatant. Connexins that integrated into the microsomes formed homo- and heterooligomeric structures. The assembled proteins in the microsome were fused with a synthetic membrane and single channel conductance was measured and found to be very similar to wild-type membranes. (1618)

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J. Biol. Chem. 272, 20756-20763. Characterization of point mutations in patients with X-linked ichthyosis. Effects On the structure and function of the steroid sulfatase protein. 1997

Alperin, E. S. and Shapiro, L. J.

Notes: Transcripts were produced using the RiboMAX™ Large Scale RNA Production System and expressed in the Rabbit Reticulocyte Lysate System with or without Canine Pancreatic Microsomal Membranes (CMMs). Proteinase K protection assays were performed with proteins expressed in the presence of the membranes with and without Triton® X-100 solubilization of the membranes. (1509)

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J. Biol. Chem. 272(33), 20756-20763. Characterization of point mutations in patients with X-linked ichthyosis: Effects on the structure and function of the steroid sulfatase protein. 1997

Alperin, E.S. and Shapiro, L.J.

Notes: The ribonuclease inhibitor was used to protect RNA during the reverse transcriptase reaction for RT-PCR. The RNasin® Ribonuclease Inhibitor is also used in the RiboMAX® System for RNA stabilization during in vitro transcription. Transcripts were produce with the RiboMAX® System and expresed in Rabbit Lysate with or without Canine Microsomal Membranes. Proteinase K protection assays are performed with proteins expressed in the presence of the membranes with and without Triton X-100 solubilization of the membranes. The proteinase K digests were stopped by the addition of PMSF and immediately loading the sample into 100°C SDS Sample buffer. (1670)

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J. Biol. Chem. 272, 31738-31746. Cig30, a mouse member of a novel membrane protein gene family, is involved in the recruitment of brown adipose tissue 1997

Tvrdik, P., Asadi, A., Kozak, L.P., Nedergaard, J., Cannon, B., Jacobsson, A.

Notes: The pCI-neo Mammalian Expression Vector was used to construct a chimeric vector with the neo resistance gene downstream of an internal ribosome entry site, so that the neo gene was transcribed with the Cig30 cDNA and both protein controlled by the CMV promoter. The construct was used to make stable transfectants of the HIB-1B hibernoma cells. Cell lines were analyzed by Northern blotting. The protein was also analyzed in vitro with the Rabbit Reticulocyte Lysate and Canine Pancreatic Microsomal Membranes (CMMs). (0246)

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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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J. Neurosci. 17, 3038-3051. ENC-1: a novel mammalian kelch-related gene specifically expressed in the nervous system encodes an actin-binding protein. 1997

Hernandez, M.C., Andres-Barquin, P.J., Martinez, S., Bulfone, A., Rubenstein, J.L., Israel, M.A.

Notes: ENC-1 and a T7 tagged ENC-1 were translated in the Rabbit Reticulocyte Lysate System to confirm the size of the protein products. (1039)

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Cell 88, 757-766. Frxb, an execreted protein expressed in the Spemann Organizer, binds and inhibits Wnt-8. 1997

Wang. S., Krinks, M., Lin, K., Luyten, F.P. and Moos, M., Jr

Notes: To demonstrate an interaction between Frzb and Wnt proteins in vitro, the proteins were cotranslated using Rabbit Reticulocyte Lysate in the presence of Canine Microsomal Membranes then immunoprecipitated with specific antibodies to either one. Control immunoprecipitations were performed with cotranslations with Frzb and the β-lactamase control or Wnt and the β-lactamase control. The procedure for the immunoprecipitation is referenced. (1603)

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J. Cell Biol. 139, 895-905. Functional expression cloning and characterization of SFT, a stimulator of Fe transport. 1997

Gutierrez, J.A., Yu, J., Rivera, S., Wessling-Resnick, M.

Notes: The cRNA for the Stimulator of Fe Transport (SFT) was in vitro translated with Rabbit Reticulocyte Lysate in the presence of Canine Microsomal Membranes. The RRL was used at 70% of total volume and 2 equivalents of the membranes were used. After translation, the reaction was treated with RNase A to reduce background. The SFT protein is predicted to have six transmembrane domains. (1621)

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J. Biol. Chem. 272, 8019-8025. In vitro reconstitution of assembly of apolipoprotein B48-containing lipoproteins. 1997

Rusiñol, A.E., Jamil, H. and Vance, J.E.

Notes: The authors prepared their own rat liver microsomes. Proteins were translated using Promega's Rabbit Reticulocyte Lysate in the presence or absence of the microsomes. The translated proteins were assayed for proteinase K protection in the presence or absence of Triton X-100. The proteinase K digests were stopped by the addition of PMSF. (1595)

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J. Virol. 71, 6373-6380. In vitro study of the NS2-3 protease of hepatitis C virus. 1997

Pieroni, L., Santolini, E., Fipaldini, C., Pacini, L., Migliaccio, G., La Monica, N.

Notes: This article describes an in vitro assay to activate hepatitis C virus (HCV) NS2-3 protease posttranslationally and methods studying the effects of several common inhibitors on enzymatic activity. HCV N2-3 protease was expressed using the Rabbit Reticulocyte Lysate System in the presence of [35S]methionine, and the NS2-3 protease was activated by detergent cleavage. The protein was incubated with various proteinase inhibitors (aprotinin, antipain, DTT, E64, pepstatin, EGTA, EDTA, iodoacetamide, N-ethylaleimide, 1,10-phenanthroline, phenylmethylsulfonyl fluoride, soybean trypsin inhibitor, TLCK and TPCK). (0533)

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Proc. Natl. Acad. Sci. USA 94, 14066-14071. Large conductance voltage- and calcium-dependent K+ channel, a distinct member of voltage-dependent ion channels with sever N-terminal tranmembrane segments (S0-S6), an extracellular N terminus, and an intracellular (S9-S10) C terminus. 1997

Meera, P., Wallner, M., Song, M. and Toro, L.

Notes: Different truncations of the protein of interest were in vitro translated in the presence of the membranes. The translation reaction were centrifuged and the pellet and supernatants assays for the presence of the protein product. (1628)

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J. Biol. Chem. 272, 18325-18332. Mapping the Ends of Transmembrane Segments in a Polytopic Membrane Protein. Scanning n-glycosylation mutagenesis of extracytosolic loops in the anion exchanger, band 3 1997

Popov, M., Tam, L.Y., Li, J., Reithmeier, R.A.F.

Notes: The authors used Promega's Canine Pancreatic Microsomal Membranes (CMMs) in combination Promega's Rabbit Reticulocyte Lysate to study the effects of various N-linked glycosylation mutations for TM mapping. Researchers included the detergent octaethylene glycol mono n-dodecyl ether from Nikkol in reactions without the CMMs to prevent aggregation of the proteins. (0541)

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J. Neurosci. 17, 8201-8212. Neuronal alpha-bungarotoxin receptors differ structurally from other nicotinic acetylcholine receptors. 1997

Rangwala, F., Drisdel, R.C., Rakhilin, S., Ko, E., Atluri, P., Harkins, A.B., Fox, A.P., Salman, S.B. and Green, W.N.

Notes: The two products were used to show that the rat a7 cDNA was translated and could be processed in vitro to a higher molecular weight, most likely by glycosylation. (1630)

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J. Biol. Chem. 272, 22097-22102. Ricin A chain can transport unfolded dihydrofolate reductase into the cytosol. 1997

Beaumelle, B., Taupiac, M.P., Lord, J.M. and Roberts, L.M.

Notes: The SILVER SEQUENCE™ DNA Sequencing System, Rabbit Reticulocyte Lysate System, Wizard® Plus Minipreps DNA Purification System and Taq DNA polymerase were used in this study. (1473)

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Proc. Natl. Acad. Sci. USA 94, 12297-12302. RNA-peptide fusions for the in vitro selection of peptides and proteins. 1997

Roberts, R.W., Szostak, J.W.

Notes: Covalent fusions of mRNA and the peptide or protein that it encodes were generated by in vitro translation in Rabbit Reticulocyte Lysate of mRNAs carrying puromycin. Puromycin, which is a peptidyl acceptor antibiotic, mimics the aminoacyl end of tRNA, enters the ribosomal A site and accepts the nascent peptide. Specific fusions of stable mRNA-peptide fusions were then selected from a pool of random sequence mRNA-peptide fusions by immunoprecipitation. This technique suggests a route for selection and evolution of proteins. (0484)

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Biochemistry 36, 11437-11443. Role of ribosomes in reinitiation of membrane insertion of internal transmembrane segments in a polytopic membrane protein. 1997

Wang, C., Chen, M., Han, E., Zhang, J. T.

Notes: The authors used the Rabbit Reticulocyte Lysate together with Canine Pancreatic Microsomal Membranes (CMMs) to study the insertion of internal transmembrane domains. They removed ribosomes from Rabbit Reticulocyte Lysate by centrifugation and replaced with ribosomes isolated from a Wheat Germ Extract and restored functional translation with resulting Rabbit Reticulocyte Lysate supernatant (ribosome-free) fraction. (0196)

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J. Biol. Chem. 272, 22191-22198. SV40 large tumor antigen nuclear import is regulated by the double- stranded DNA-dependent protein kinase site (Serine 120) flanking the nuclear localization sequence 1997

Xiao, C. Y. , Hubner, S. , Jans, D. A.

Notes: Promega's Casein Kinase II and DNA-Dependent Protein Kinase were used for in vitro phosphorylation of the SV40 large T antigen expressed either from Rabbit Reticulocyte Lysate or a large Tantigen-β-Gal fusion protein expressed in a rat hepatoma cell line. Lysates and cell extracts were assayed for DNA-dependent protein kinase activity with a peptide substrate. The ability of β-Gal-T antigen fusions (± phosphorylation) to bind to murine importin subunits was assessed by an ELISA system using the Anti-β-Galactosidase mAb. (0120)

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J. Biol. Chem. 272, 20936-20944. The p38/Reactivating kinase mitogen-activated protein kinase cascade mediates the activation of the transcription factor insulin upstream factor 1 and insulin gene transcription by high glucose in pancreatic beta-cells. 1997

Macfarlane, W.M., Smith, S.B., James, R.F.L., Clifton, A.D., Doza, Y.N., Cohen, P., Docherty, K.

Notes: The AMV Reverse Transcriptase, RNasin® Ribonuclease Inhibitor and Taq DNA Polymerase were used for RT-PCR to isolate a clone of the human insulin upstream factor 1, a transcription factor. The protein was expressed in vitro with the Rabbit Reticulocyte Lysate and used for gel shift assays. Reporter studies were performed in MIN6 cells. (0762)

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J. Biol. Chem. 272, 6119-6127. Topological rules for membrane protein assembly in eukaryotic cells. 1997

Gafvelin, G., Sakaguchi, M., Andersson, H. and von Heijne, G.

Notes: The paper looked at the insertion of a model protein with one, two and four transmembrane segments and different distributions of positively charged residues in the N-terminal tail and the polar loops. The proteins were translated in the presence of the microsomes. Translocation of polypeptides to the lumenal side of the microsomes was assayed by prevention of N-linked glycosylation through competitive inhibition by the addition of a glycosylation acceptor tripeptide but not by a nonacceptor tripeptide and by proteinase K treatment of the microsomes. All techniques are referenced. (2033)

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