Human TGFB1 Gene in Lentivector, Puromycin-selectable, Endotoxin-free DNA
The TGFβ (transforming growth factor β) superfamily proteins are pleiotropic cytokines that regulate a diverse range of cellular processes, including proliferation, differentiation, migration, adhesion and apoptosis. The TGFβ superfamily elicits 4 signaling pathways: TGFβ, Bone Morphogenic Protein (BMP), Activin, and Nodal. Each pathway signals through a heteromeric receptor complex composed of at least 2 type-I and 2 type-II receptors with cytoplasmic serine/threonine kinase domains. The receptor complex phosphorylates and activates Smad proteins to regulate downstream gene transcription. There are 3 closely related mammalian members (TGFβ1-3) of the TGFβ family that share a characteristic cystine knot structure. They are known to function as cellular switches that regulate immune function, proliferation and epithelialmesenchymal transition (EMT). Each TGFβ isoform has non-redundant functions. For instance, TGFβ1-deficient mice display defects in hematopoiesis and endothelial differentiation, and die of overwhelming inflammation. TGFβ1 is highly expressed in bone and acts as a stimulator of bone formation. A furinlike convertase processes it to generate an Nterminal 249 aa latencyassociated peptide (LAP) and a Cterminal 112 aa mature TGFβ1. Mature human TGFβ1 is 100% identical to pig, dog and cow TGFβ1, and 99% identical to mouse, rat and horse TGFβ1, therefore showing crossspecies activity. TGFβs are identified as potential therapeutic targets for cancer and cardiovascular diseases. This gene is frequently upregulated in tumor cells, and mutations in this gene result in Camurati-Engelmann disease.
Gene Symbol: TGF-beta1; CED; LAP; DPD1; TGFB; TGFbeta
NCBI Gene ID: 7040
Uniprot Entry: P01137
Construct Details: Full length human TGFB1 gene is subcloned into the Lentiviral expression vector pLTC with an upstream CMV promoter, which can be used for both transient and stable expression in mammalian cells. The construct also contains a Puromycin marker for antibiotic stable selection. It can be co-transfected with the LentiPAK DNA mix (SKU# LP-001) into HEK293 cells to produce high titer lentiviral particles.
Vector Type: pLTC-IRES-Puro (lentiviral expression vector containing a heterologous CMV promoter and a puromycin-selectable marker, see the vector map above)
Formulation: Supplied with 10 μg of endotoxin-free plasmid DNA at 200 ng/μl in sterile TE buffer (pH8.0)
Gene Insert Size: 1173 (bp)
Gene Insert Sequence:
ATGCCGCCCTCCGGGCTGCGGCTGCTGCCGCTGCTGCTACCGCTGCTGTGGCTACTGGTGCTGACGCCTGGCCGGCCGGC
CGCGGGACTATCCACCTGCAAGACTATCGACATGGAGCTGGTGAAGCGGAAGCGCATCGAGGCCATCCGCGGCCAGATCC
TGTCCAAGCTGCGGCTCGCCAGCCCCCCGAGCCAGGGGGAGGTGCCGCCCGGCCCGCTGCCCGAGGCCGTGCTCGCCCTG
TACAACAGCACCCGCGACCGGGTGGCCGGGGAGAGTGCAGAACCGGAGCCCGAGCCTGAGGCCGACTACTACGCCAAGGA
GGTCACCCGCGTGCTAATGGTGGAAACCCACAACGAAATCTATGACAAGTTCAAGCAGAGTACACACAGCATATATATGT
TCTTCAACACATCAGAGCTCCGAGAAGCGGTACCTGAACCCGTGTTGCTCTCCCGGGCAGAGCTGCGTCTGCTGAGGCTC
AAGTTAAAAGTGGAGCAGCACGTGGAGCTGTACCAGAAATACAGCAACAATTCCTGGCGATACCTCAGCAACCGGCTGCT
GGCACCCAGCGACTCGCCAGAGTGGTTATCTTTTGATGTCACCGGAGTTGTGCGGCAGTGGTTGAGCCGTGGAGGGGAAA
TTGAGGGCTTTCGCCTTAGCGCCCACTGCTCCTGTGACAGCAGGGATAACACACTGCAAGTGGACATCAACGGGTTCACT
ACCGGCCGCCGAGGTGACCTGGCCACCATTCATGGCATGAACCGGCCTTTCCTGCTTCTCATGGCCACCCCGCTGGAGAG
GGCCCAGCATCTGCAAAGCTCCCGGCACCGCCGAGCCCTGGACACCAACTATTGCTTCAGCTCCACGGAGAAGAACTGCT
GCGTGCGGCAGCTGTACATTGACTTCCGCAAGGACCTCGGCTGGAAGTGGATCCACGAGCCCAAGGGCTACCATGCCAAC
TTCTGCCTCGGGCCCTGCCCCTACATTTGGAGCCTGGACACGCAGTACAGCAAGGTCCTGGCCCTGTACAACCAGCATAA
CCCGGGCGCCTCGGCGGCGCCGTGCTGCGTGCCGCAGGCGCTGGAGCCGCTGCCCATCGTGTACTACGTGGGCCGCAAGC
CCAAGGTGGAGCAGCTGTCCAACATGATCGTGCGCTCCTGCAAGTGCAGCTGA
Primers: Gene insert coding sequence can be confirmed by following primers:
(a) forward (or 5'-end) primer: 5’-CAAATGGGCGGTAGGCGTG-3’;
(b) reverse (or 3'-end) primer: 5’-CCAGGTTTCCGGGCCCTCAC-3’
Note: The sequence may vary due to the naturally occurring variants or mutations, such as single nucleotide polymorphism, or the artifact during PCR amplification. High fidelity PCR systems should always be used.
FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.
Restriction: This product is not transferable or re-sellable. Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose. Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules. Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction. Your use of this product constitutes acceptance of the terms of this limited use agreement. Please refer to our “terms & conditions” for details. If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.
The product is shipped at 4°C. Upon receipt, centrifuge the product briefly before opening the vial. It is recommended to store small aliquots at the temperature below –20°C for long-term storage and the product is stable for 6 months.
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.
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5. Cytokine Growth Factor Rev. 18: 279 (2007).
Product datasheet (pdf) can be downloaded here: LTP2092P-PDS.pdf
Additional supporting documents, including COA and MSDS are available upon request.
FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.
Restriction: This product is not transferable or re-sellable. Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose. Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules. Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction. Your use of this product constitutes acceptance of the terms of this limited use agreement. Please refer to our “terms & conditions” for details. If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.