Human CD73/NT5E/NTE/5’-NT Gene, Full-length cDNA Clone in Lentivector, Endotoxin-free DNA

Description

CD73, also known as 5’-NT, NT5 and NTE, is a glycosyl-phosphatidyl-inositol (GPI) anchored cell membrane protein that belongs to the 5'-nucleotidase (5’-NT) family. The native enzyme exists as a homodimer on the cell surface. CD73 is a purine salvage enzyme expressed on human T and B lymphocytes. CD73 converts extracellular nucleoside­5'­monophosphates to membrane permeable nucleosides, with AMP as the preferred substrate. It also exhibits AMP-, NAD-, and NMN-nucleosidase activities.  CD73 is one of several ectoenzymes responsible for the production of extracellular adenosine, a signaling molecule involved in many physiologic processes, including inflammation, tissue injury response, and cholinergic synapses. CD73 produces extracellular adenosine, which then acts on G protein-coupled purigenic receptors (GPCR) to induce intracellular responses. CD73 is also a regulator of leukocyte extravasation, a function that requires its 5'­nucleotidase activity. CD73 mediates lymphocyte-stromal cell interactions to facilitate lymphocyte development/function.  CD73 has also been considered as a lymphocyte maturation marker independent of its catalytic activity. CD73 serves as a co-stimulatory molecule in activating T cells. The mutation in CD73 gene is associated with calcification of joints and arteries (CALJA), a condition characterized by adult-onset calcification of the lower extremity arteries  and hand and foot capsule joints.

 

 

Gene Symbol: CD73; 5’-NT; NT; eN; NT5; NTE; eNT; E5NT; CALJA

 

NCBI Gene ID: 4907

 

Uniprot Entry: P21589

 

Construct Details: Full length human CD73 gene is subcloned into the Lentiviral expression vector pLTC with an upstream CMV promoter and a selection marker, which can be used for both transient and stable expression in mammalian cells.  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 or pLTC-IRES-Marker (lentiviral expression vector containing a heterologous CMV promoter, 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: 1725 (bp)

 

Gene Insert Sequence:  

ATGTGTCCCCGAGCCGCGCGGGCGCCCGCGACGCTACTCCTCGCCCTGGGCGCGGTGCTGTGGCCTGCGG

CTGGCGCCTGGGAGCTTACGATTTTGCACACCAACGACGTGCACAGCCGGCTGGAGCAGACCAGCGAGGA

CTCCAGCAAGTGCGTCAACGCCAGCCGCTGCATGGGTGGCGTGGCTCGGCTCTTCACCAAGGTTCAGCAG

ATCCGCCGCGCCGAACCCAACGTGCTGCTGCTGGACGCCGGCGACCAGTACCAGGGCACTATCTGGTTCA

CCGTGTACAAGGGCGCCGAGGTGGCGCACTTCATGAACGCCCTGCGCTACGATGCCATGGCACTGGGAAA

TCATGAATTTGATAATGGTGTGGAAGGACTGATCGAGCCACTCCTCAAAGAGGCCAAATTTCCAATTCTG

AGTGCAAACATTAAAGCAAAGGGGCCACTAGCATCTCAAATATCAGGACTTTATTTGCCATATAAAGTTC

TTCCTGTTGGTGATGAAGTTGTGGGAATCGTTGGATACACTTCCAAAGAAACCCCTTTTCTCTCAAATCC

AGGGACAAATTTAGTGTTTGAAGATGAAATCACTGCATTACAACCTGAAGTAGATAAGTTAAAAACTCTA

AATGTGAACAAAATTATTGCACTGGGACATTCGGGTTTTGAAATGGATAAACTCATCGCTCAGAAAGTGA

GGGGTGTGGACGTCGTGGTGGGAGGACACTCCAACACATTTCTTTACACAGGCAATCCACCTTCCAAAGA

GGTGCCTGCTGGGAAGTACCCATTCATAGTCACTTCTGATGATGGGCGGAAGGTTCCTGTAGTCCAGGCC

TATGCTTTTGGCAAATACCTAGGCTATCTGAAGATCGAGTTTGATGAAAGAGGAAACGTCATCTCTTCCC

ATGGAAATCCCATTCTTCTAAACAGCAGCATTCCTGAAGATCCAAGCATAAAAGCAGACATTAACAAATG

GAGGATAAAATTGGATAATTATTCTACCCAGGAATTAGGGAAAACAATTGTCTATCTGGATGGCTCCTCT

CAATCATGCCGCTTTAGAGAATGCAACATGGGCAACCTGATTTGTGATGCAATGATTAACAACAACCTGA

GACACACGGATGAAATGTTCTGGAACCACGTATCCATGTGCATTTTAAATGGAGGTGGTATCCGGTCGCC

CATTGATGAACGCAACAATGGCACAATTACCTGGGAGAACCTGGCTGCTGTATTGCCCTTTGGAGGCACA

TTTGACCTAGTCCAGTTAAAAGGTTCCACCCTGAAGAAGGCCTTTGAGCATAGCGTGCACCGCTACGGCC

AGTCCACTGGAGAGTTCCTGCAGGTGGGCGGAATCCATGTGGTGTATGATCTTTCCCGAAAACCTGGAGA

CAGAGTAGTCAAATTAGATGTTCTTTGCACCAAGTGTCGAGTGCCCAGTTATGACCCTCTCAAAATGGAC

GAGGTATATAAGGTGATCCTCCCAAACTTCCTGGCCAATGGTGGAGATGGGTTCCAGATGATAAAAGATG

AATTATTAAGACATGACTCTGGTGACCAAGATATCAACGTGGTTTCTACATATATCTCCAAAATGAAAGT

AATTTATCCAGCAGTTGAAGGTCGGATCAAGTTTTCCACAGGAAGTCACTGCCATGGAAGCTTTTCTTTA

ATATTTCTTTCACTTTGGGCAGTGATCTTTGTTTTATACCAATAG

 

 

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.

 

 

1.  Cell Signal. 9: 131 (1997)

2. Immunol. Rev. 161:95 (1998).

3. Eur J Immunol. 28: 2981 (1998)

4. J. Biol. Chem. 278:13468 (2003)

5. J Immunol. 180: 4246 (2008)

 

 

 

 

Product datasheet (pdf) can be downloaded here: LTP2157-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.