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Biol. Reprod. 79, 938–946. Japanese eel follicle-stimulating hormone (Fsh) and luteinizing hormone (Lh): production of biologically active recombinant Fsh and Lh by Drosophila S2 cells and their differential actions on the reproductive biology. 2008

Kazeto, Y., Kohara, M., Miura, T., Miura, C., Yamaguchi, S., Trant, J.M., Adachi, S. and Yamauchi, K.

Notes: The ORFs of Japanese eel follicle-stimulating hormone receptor (Fshr) and luteinizing hormone receptor (Lhr) were ligated into the pSI Mammalian Expression Vector. Fifty nanograms of this construct or the pSI Vector was cotransfected with 0.5ng of pRL-null Vector and 1µg or a cAMP-responsive firefly luciferase reporter vector into COS cells seeded in 60mm dishes. The next day, the cells were trypsinized, replated into 96-well plates and allowed to recover for 1 day. The transfected cells were serum-starved then treated with hormones six hours before being lysed and enzymatic activity measured with the Dual-Luciferase® Reporter Assay System. (3988)

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J. Biol. Chem. 282, 5506-5513. HERG is protected from pharmacological block by alpha-1,2-glucosyltransferase function. 2007

Nakajima, T., Hayashi, K., Viswanathan, P.C., Kim, M.Y., Anghelescu, M., Barksdale, K.A., Shuai, W., Balser, J.R. and Kupershmidt, S.

Notes: The pSI Mammalian Expression Vector was used to subclone HERG (human ether-à-go-go-related gene) cDNA and then subjected to site-directed mutagenesis. The constructs were used for transient transfection and tested for membrane current. (3756)

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Hum. Mol. Genet. 15, 999–1013. An exon skipping-associated nonsense mutation in the dystrophin gene uncovers a complex interplay between multiple antagonistic splicing elements. 2006

Disset, A., Bourgeois, C.F., Benmalek, N., Claustres, M., Stevenin, J. and Tuffery-Giraud, S.

Notes: To construct dystrophin minigenes, genomic DNA containing a mutation in dystrophin was amplified for exons 30, 31 and 32. The three PCR fragments were combined and amplified into one product. This overlap-extension PCR generated two minigenes which were then cloned into the pGEM®-T Vector and sequenced. After EcoR I digestion, the minigenes were ligated into the pSI Mammalian Expression Vector and transiently transfected into C2C12 cells for expression analysis. (3499)

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Br. J. Pharmacol. 147, 73–82. Characterization of the Mas-related gene family: structural and functional conservation of human and rhesus MrgX receptors. 2006

Burstein, E.S., Ott, T.R., Feddock, M., Ma, J.N., Fuhs, S., Wong, S., Schiffer, H.H., Brann, M.R. and Nash, N.R.

Notes: This study examined both the human and the macaque counterparts of the Mas-related genes (Mrg), a family of G-protein-coupled receptors, in assays to compare function and structure. Clones of adenylyl cyclase type II (AC2) and Gαo were subcloned into the pSI Mammalian Expression Vector and used in the co-transfection experiments with the Mrgs. Receptor Selection and Amplification Technology (R-SAT™) was used for this comparison of the human and macaque Mrgs. Cells were plated in 96-well plates and transiently transfected with 57ng of 4 different plasmids including one of the Mrg receptors, 2ng AC2 and 30ng pSV-β-galactosidase Vector. One day post-transfection, ligands were added and after 5 days, the β-galactosidase expression was assessed. (3551)

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J. Immunol. 174, 4647–4656. Translation from cryptic reading frames of DNA vaccines generates an extended repertoire of immunogenic, MHC class I-restricted epitopes. 2005

Schirmbeck, R., Riedl, P., Fissolo, N., Lemonnier, F.A., Bertoletti, A. and Reimann, J.

Notes: To analyze whether CD8+ T cell responses can be generated from gene products translated from another reading frame from the primary ORF encoded by DNA vaccines, various constructs were made using the pCI Mammalian Expression Vector. These inserts included the small hepatitis B surface antigen (HBsAg) and its 3’ untranslated sequence up to the hepatitis B virus (HBV) poly(A) sequence, which contains the sequence encoding the C-terminal Pol344–832 fragment; the 832 residue Pol protein, and the large HBsAg in an alternative reading frame and an unrelated in frame and out of frame OVA OVA18–385 fragment. In the case of the OVA fragment, the CMV promoter was removed and substituted with the SV40 promoter from pSI Mammalian Expression Vector. The immune response to both the primary and secondary gene products was determined using transient transfection of LMH cells followed by Western blotting or immunoprecipitation or counting CD8+ T cells from spleen cells. (3553)

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Cancer Res. 62, 5668 – 5671. A novel mechanism of nuclear factor κB activation through the binding between inhibitor of nuclear factor-κBα  and the processed NH2-terminal region of Mig-6. 2002

Tsunoda, T., Inokuchi, J., Baba, I., Okumura, K., Naito, S., Sasazuki, T., and Shirasawa, S.

Notes: Mig-6 cDNA was cloned into the pSI Vector to be used in cotransfection experiments with luciferase reporter constructs.  The phRL-CMV Vector was added to the transfections to normalize the data generated by the Dual-Luciferase® Reporter Assay System in NIH/3T3 cells. (2632)

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J. Biol. Chem. 274, 21228-21233. Human mitochondrial carbonic anhydrase VB: cDNA cloning, mRNA expression, subcellular localization, and mapping to chromosome X. 1999

Fujikawa-Adachi, K., Nishimori, I., Taguchi, T., Onishi, S.

Notes: The two carbonic anhydrase isoforms were cloned into the pSI Mammalian Expression Vector. The proteins were each transiently expressed in COS-7 cells. (1128)

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J. Biol. Chem. 274, 18382-18386. The agonist-binding domain of the calcium-sensing receptor is located at the amino-terminal domain. 1999

Brauner-Osborne, H., Jensen, A.A., Sheppard, P.O., O'Hara, P., Krogsgaard-Larsen, P.

Notes: Both the calcium-sensing receptor and the metabotropic glutamate receptor 1 were each subcloned into the pSI Mammalian Expression Vector. The estimated molecular weight of the two proteins are 56kDa and 55kDa, respectively. The proteins were transiently expressed in tsA cells, a transformed HEK 293 cell line. (1402)

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J. Biol. Chem. 273, 27231-27235. A K+ channel splice variant common in human heart lacks a c-terminal domain required for expression of rapidly activating delayed rectifier current. 1998

Kupershmidt, S., Snyders, D.J., Raes, A., Roden, D.M.

Notes: The pSI Mammalian Expression Vector was used to express the 127kDa human ether-a-go-go-related gene (HERG) cDNA and mutants in Ltk- cells. The expression vectors were cotransfected with a green fluorescent protein vector to assess transfection efficiency and to select transfected cells by FACS for patch/clamp studies. (0865)

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J. Biochem. 273, 34527-34534. Identification of the half-cystine residues in porcine submaxillary mucin critical for multimerization through the D-domains. Roles of the CGLCG motif in the D1- and D3-domains. 1998

Perez-Vilar, J., Hill, R.L.

Notes: The authors used Promega's pSI Mammalian Expression Vector to express protein in COS-7 cells. (0559)

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