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Promega Corporation

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Oncogene 20, 1041-1051. Isolation and characterization of the TERE1 gene, a gene down-regulated in transitional cell carcinoma of the bladder 2001

McGarvey, T.W., Nguyen, T., Tomaszewski, J.E., Monson, F.C. and Malkowicz, S.B.

Notes: The cDNA for the TERE1 gene was amplified by PCR and directly subcloned into the pTARGET™ Mammalian Expression Vector. The 338-amino acid (36.8kDa) TERE1 protein was stably expressed in three human bladder transitional cell carcinomas (J82, T24 and 1376 TCC) and the human embryonic kidney cell line, HEK 293. Stable transfectants were selected with the antibiotic G-418. Overexpression of the protein caused 80-90% inhibition of cellular proliferation in two of the transitional cell carcinoma cell lines and inhibition of aneuploidy in the third. Transfections were accomplished in 24-well plates with the Tfx™-20 Transfection Reagent. Excellent details of the 1 hour transfection protocol are presented. (2596)

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J. Bacteriol. 183, 7094-7101. Quantification of expression of Staphylococcus epidermidis housekeeping genes with Taqman quantitative PCR during in vitro growth and under different conditions. 2001

Vandecasteele, S.J., Peetermans, W.E., Merckx, R. and Van Eldere, J.

Notes: M-MLV Reverse Transcriptase and RNasin® Ribonuclease Inhibitor were used in a reaction to generate first strand cDNA.  The generated cDNA was used as a template for quantitative PCR in a Taqman® Assay.  The pGEM®-T Vector System was used to clone copies of each target gene so that standards could be generated.  The Wizard® Genomic DNA Purification Kit was used to isolated genomic DNA from the S. epidermidis to generate a standard curve for quantitation of genomic DNA contamination of samples. (2291)

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Brain Res. Mol. Brain Res. 76, 25–35. Expression of the GDNF family members and their receptors in the mature rat cochlea. 2000

Stöver, T., Gong, T.L., Cho, Y., Altschuler, R.A. and Lomax, M.I.

Notes: Total RNA was isolated from various rat tissues with the SV Total RNA Isolation System. Yields are reported as 10µg from 16 whole cochlea, 8µg from 16 modiola, 10.4µg from 48 cochlear sensorineural epithelial/lateral walls and 50µg from the substantia nigra region of four brains. The isolated RNA was used for RT-PCR in the presence of RNasin® Ribonuclease Inhibitor. The resulting amplimer was subcloned into the pGEM®-T Easy Vector and clones were purified with the Wizard® Plus SV Minipreps System. (2176)

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Nat. Cell Biol. 2, 757-761. Mammalian recombination-repair genes XRCC2 and XRCC3 promote correct chromosome segregation. 2000

Griffin, C.S., Simpson, P.J., Wilson, C.R., and Thacker, J.

Notes: The XRCC3 gene was amplified from a human testis cDNA library and directly subcloned into the pTARGET™ Mammalian Expression Vector. The insert was sequenced, then electroporated into the irs1SF hamster radiation-sensitive cell line. Stable transfectants were isolated by G-418 selection and used in chromosome segregation studies. (2599)

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J. Immunol. 164, 2248-2254. Successful TCR-based immunotherapy for autoimmune myocarditis with DNA vaccines after rapid identification of pathogenic TCR. 2000

Matsumoto, Y., Jee, Y. and Sugisaki, M.

Notes: The T-cell receptor of interest was amplified and subcloned in the pTARGET™ Mammalian Expression Vector. Large amounts of the vector were purified with the Wizard® Plus MegaPreps DNA Purification System and used as a DNA vaccine in Lewis rats. (0701)

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J. Immunol. 164, 3961-3970. Differential localization and regulation of death and decoy receptors for TNF-related apoptosis-inducing Ligand (TRAIL) in human melanoma cells 2000

Zhang, X.D., Franco, A.V., Nguyen, T., Gray, C.P., Hersey, P.

Notes: The pTARGET™ Mammalian Expression Vector was used to clone and express the TRAIL-3 and the TRAIL-4 receptors. The proteins were expressed in primary human melanoma cells. (0098)

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J. Biol. Chem. 275, 11666-11671. Distinct roles for recombinant cytosolic 5'-nucleotidase-I and -II in AMP and IMP catabolism in COS-7 and H9c2 rat myoblast cell lines 2000

Sala-Newby, G.B., Freeman, N.V.E., Skladanowski, A.C., Newby, A.C.

Notes: The pTargeT™ Mammalian Expression Vector System was used to clone and express the human cytosolic 5'-nucleotidase II, a 65kDa protein. The protein was expressed and assayed in COS-7 monkey kidney cells and H9c2 rat myoblast cells. The pSV-beta Galactosidase Control Vector was used as a control for transfection efficiency. (0431)

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J. Biol. Chem. 274, 8391-8404. Cyclic AMP- and Cyclic GMP-dependent protein kinases differ in their regulation of cyclic AMP response element-dependent gene transcription. 1999

Collins, S.P., Uhler, M.D.

Notes: The pSP73 Vector was used for routine subcloning. The pGEM®-T Vector System was used for subcloning of PCR products. (1320)

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J. Biol. Chem. 274, 2271-2278. Antioxidant function of the mitochondrial protein SP-22 in the cardiovascular system. 1999

Araki, M., Nanri, H., Ejima, K., Murasato, Y., Fujiwara, T., Nakashima, Y. and Ikeda, M.

Notes: The authors used the Tfx™-50 Reagent to transfect bovine aortic endothelial cells. They also used the pGEM®-T Easy Vector System to clone PCR products. (1482)

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J. Biol. Chem. 274, 7982-7986. KSR-1 binds to G-protein beta gamma subunits and inhibits beta gamma-induced mitogen-activated protein kinase activation. 1999

Bell, B., Xing, H., Yan, K., Gautam, N. and Muslin, A.J.

Notes: The 873 amino acid kinase repressor of Ras (KSR-1) was amplified to add a flag tag and the resultant product was cloned into the the pTARGET™ Mammalian Expression Vector. The protein was expressed in COS7 cells and used for subcellular fractionation studies. (1434)

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J. Immunol. 162, 871-877. Alternative splicing and hypermutation of a nonproductively rearranged TCR alpha-chain in a T cell hybridoma. 1999

Marshall, B., Schulz, R., Zhou, M., Mellor, A.

Notes: The Access RT-PCR System was used to amplify Vα1024 mRNA of the TCR alpha chain and the resulting product was cloned with the pGEM®-T Vector System. (0734)

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J. Biol. Chem. 274, 24865-24872. Nuclear localization of protein kinase U-alpha is regulated by 14-3-3. 1999

Zhang, S., Xing, H., Muslin, A.J.

Notes: The protein named KIAA (identified by yeast two-hybrid screen) and the 14-3-3zeta protein were each amplified to add epitope tags. The resulting products were subcloned into the pTARGET™ Mammalian Expression Vector and stably expressed in NIH/3T3 cells. (0096)

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Genetics 153, 1743-1751. Insights into genome differentiation: pheromone-binding protein variation and population history in the European corn borer (Ostrinia nubilalis). 1999

Willett, C. S., Harrison, R. G.

Notes: In this paper, DNA visualized on a sequencing gel is stained with the SILVER SEQUENCE™ DNA Sequencing Systems, and PCR products are cloned into the pGEM®-T Vector. (0187)

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Am. J. Hum. Genet. 65, 1040-1046. Temperature-Sensitive RB Mutations Linked to Incomplete Penetrance of Familial Retinoblastoma in 12 Families. 1999

Otterson, G.A., Modi, S., Nguyen, K., Coxon, A.B., Kaye, F.J.

Notes: A PCR product amplified from a mutant RB (retinoblastoma protein) cDNA was cloned into the pCI-neo Mammalian Expression Vector. In vivo cyclin-mediated phosphorylation was assayed by cotransfecting the RB expression plasmids with members of the cyclin D or cyclin E family into an RB (-/-) cell line. (0571)

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J. Biol. Chem. 274, 17789-17793. The mechanism of adenosine formation in cells: Cloning of cytosolic 5'-nucleotidase I 1999

Sala-Newby, G.B., Skladanowski, A.C., Newby, A.C.

Notes: A clone of the cytosolic 5'-nucleotidase I cDNA into the available BamHI-KpnI site of the pTargeT™ Mammalian Expression Vector System. The expression construct was cotransfected into COS-7 cells with the pSV-betaGalactosidase Vector to control for transfection efficiency. (0432)

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J. Neurosci. 18, 16-25. Regulation of Ca2+-dependent K+ channel expression in rat cerebellum during postnatal development. 1998

Muller, Y.L., Reitstetter, R., Yool, A.J.

Notes: Poly-A+ RNA was isolated from rat cerebellar total RNA with the PolyATtract® mRNA Isolation System and used for semi-quantitative RT-PCR. The targets of the RT-PCR were cloned with the pGEM®-T Vector System and sequenced to show they quantitated the correct targets. (0671)

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Appl. Environ. Microbiol. 64, 1147-1152. Common elements regulating gene expression in temperate and lytic bacteriophages of Lactococcus species. 1998

Walker, S. A. , Dombroski, C. S. , Klaenhammer, T. R.

Notes: The pGEM®-T Easy Vector was used to clone PCR products amplified from the prophage inserts of of various lysogenic Lactococcus strains. (0193)

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Gastroenterology 114, 471-481. Abnormal intestinal intraepithelial lymphocytes in refractory sprue. 1998

Cellier, C., Patey, N., Mauvieux, L., Jabri, B., Delabesse, E., Cervoni, J., Burtin, M., Guy-Grand, D., Bouhnik, Y., Modiliani, R., Barbier, J., Macintyre, E., Brousse, N. and Cerf-Bensussan, N.

Notes: The authors used the pGEM®-T Easy Vector System in their studies. (1966)

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Infect. Immun. 66, 3470-3475. Multigene families encoding the major hemagglutinins in phylogenetically distinct mycoplasmas 1998

Noormohammadi, A.H., Markham, P.F., Duffy, M.F., Whithear, K.G., Browning, G.F.

Notes: PCR products were cloned into the PinPoint® Xa1 T-Vector and expressed in JM109 cells. The cell extracts were electrophoresed and immunoblotted for the specific protein. (0626)

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Biochemistry 37, 4420-4428.. Silent nucleotide substitution in the sterol 27-hydrosylase gene (CYP 27) leads to alternative pre-mRNA splicing by activating a crytic 5' splice site at the mutant codon in cerebrotendinous xanthomatosis patients. 1998

Chen, W., Kubota, S., Teramoto, T., Nishimura, Y., Yonemoto, K., Seyama, Y.

Notes: Genomic analysis noted a single base change between wild type and mutant. To see if this base change was enough for alternative mRNA splicing, minigenes were constructed and subcloned into the pTargeT™ Vector. The mutant and wild-type minigenes were transfected into COS-1 cells. Forty-eight hours post-transfection total RNA was isolated and analyzed by RT-PCR. Transfectants with the mutant sequence did produce a different product due to alternative splicing. (1331)

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Biochemistry 37, 15050-15056. A novel Arg362Ser mutation in the sterol 27-hydroxylase gene (CYP27): Its effects on pre-mRNA splicing and enzyme activity. 1998

Chen, W., Kubota, S., Ujike, H., Ishihara, T., Seyama, Y.

Notes: A minigene was constructed with five exons and four introns from exons 5-9 of the CYP27 gene. The 2111bp product was generated with primers containing a start codon and a stop codon. The product was directly cloned into the pTARGET™ Mammalian expression vector and transfected into COS-1cells. RNA was isolated from these cells and RT-PCR was performed to amplify the spliced product. This assay could distinguish between normal splicing of the third intron and aberrant splicing of due to the Arg362Ser mutation. The proteins produced by the normal and mutant minigenes were also assessed by immunoblotting. (1333)

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Genetics 150, 1125-1131. Mouse Brachyury the Second (T2) is a gene next to classical T and a candidate gene for tct. 1998

Rennebeck, G., Lader, E., Fujimoto, A., Lei, E.P., Artzt, K.

Notes: Streptavidin MagneSphere® Paramagnetic Particles (SA-PMPs) were used to capture biotinylated primer probe in a random access retrieval of genetic information by PCR (rargip) screening [ABE, K., (1992) Rapid isolation of desired sequences from lone linker PCR amplified cDNA mixtures: application to identification and recovery of expressed sequences in cloned genomic DNA. Mamm. Genome 2:252-259]. The pGEM®-T Vector System and PolyATract® mRNA Isolation System were also used. (0513)

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Proc. Natl. Acad. Sci. USA 94, 5273-5277. Molecular cloning, sequence, expression, and processing of the interleukin 16 precursor. 1997

Baier, M., Bannert, N., Werner, A., Lang, K. and Kurth, R.

Notes: To identify the true transcriptional start of the IL-16 mRNA methods for rapid amplification of cDNA ends were applied. The complete pro-IL-16 cDNA was cloned, sequenced, and expressed in COS-7 cells. The authors used the pGEM®-T Vector System to clone fragments that were then used as templates in the Riboprobe® in Vitro Transcription System. (1453)

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J. Biol. Chem. 272, 9464-9473. Intracellular Ca2+ pool content and signaling and expression of a calcium pump are linked to virulence in Leishmania mexicana amazonesis amastigotes. 1997

Lu, H.G., Zhong, L., Chang, K.P., Docampo, R.

Notes: PolyATtract® mRNA Isolation System was used to isolate Poly A+ RNA from the protozoan Leishmania mexicanna amazonesis total RNA. The isolated RNA was used for RT-PCR. The RT-PCR products were cloned into the pGEM®-T Vector. (0748)

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Proc. Natl. Acad. Sci. USA 94, 2415-2420. Floral homeotic genes were recruited from homologous MADS-box genes preexisting in the common ancestor of ferns and seed plants. 1997

Munster, T., Pahnke, J., Di Rosa, A., Kim, J.T., Martin, W., Saedler, H., Theissen, G.

Notes: The 3'-RACE products were cloned with the pGEM®-T Vector System. (0673)

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