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)

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)

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)

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)

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)

Notes: The Access RT-PCR System was used to amplify Vα₁₀Jα₂₄ mRNA of the TCR alpha chain and the resulting product was cloned with the pGEM^®-T Vector System. (0734)

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)

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

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)

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 pTARGET™ Mammalian Expression Vector. The protein was expressed in COS7 cells and used for subcellular fractionation studies. (1434)

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)

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)

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)

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)

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

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

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)

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)

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)

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)

Notes: This article describes the use of the TNT^® T7 Quick Coupled Transcription/Translation System in a process called the ‘ARM’ technique where antibody-ribosome-mRNA (ARM) complexes are selected by protein binding. The ARM complexes are generated during translation of mRNAs that lack stop codons. The functional antibody that is still complexed with the ribosome and mRNA is selected by protein-coupled magnetic beads, and the associated mRNA is reverse-transcribed and amplified by polymerase chain reaction (RT-PCR). The PCR products are then used in subsequent cycles of the method. (2016)

Notes: Taq DNA Polymerase was used for PCR. The PCR products were purified with the Wizard^® PCR Preps DNA Purification System and cloned with the aid of the pGEM^®-T Vector System. (0624)

Notes: RT-PCR was performed with degenerate primers and the resulting 110bp product was subcloned with the pGEM®-T Vector System. The 110bp fragment was used to screen a cDNA library and four positive plaques were identified. (0650)

Notes: Promega's pGEM®-T Vector System, pCI Mammalian Expression Vector and TNT® Coupled Reticulocyte Lysate System were used in this study. (1951)

Notes: NGF was used to induce PC12 cell differentiation 3-5 days prior to assay. PCR amplimers representing various subunits of the acetylcholine receptor were subcloned into the pGEM^®-T Vector. The resulting clones were used to produce RNA probes for RNase protection assays. Also, a chimeric a5/7-HT3 receptor was generated by PCR, subcloned into the pGEM^®-T Vector and sequenced. (1419)