Human GPC3 Full-length Gene in Lentivector, Endotoxin-free DNA

Description

GPC3 (Glypican-3) is a member of the Glypican (GPC) family. Glypicans are a family of heparan sulfate proteoglycans that are linked to the cell surface by a glycosylphosphatidylinositol (GPI) anchor. Six members of this family have been found in mammals. All of them contain an N­terminal signal peptide, a large globular cysteine­rich domain (CRD) with 14 conserved cysteine residues, a stalk­like region containing the heparan sulfate attachment sites, and a C­terminal GPI anchoring site. GPC3 is processed by convertases that generates at least 3 ploypeptides: the 1st starts with Gln25, the 2nd starts with Ser359 after a furin cleavage site, and the 3rd starts with Val483. Therefore 2-3 different MW species are generated in the recombinant GPC3 preparation. They appear to be linked by disulfide bonds. GPC3 is involved in the suppression/modulation of growth in the predominantly mesodermal tissues and organs. GPC3 may regulate growth and tumor predisposition. Mutations in GPC3 cause a rare disorder Simpson­Golabi­Behmel Syndrome, which is characterized by pre- and post-natal overgrowth of multiple tissues and organs and an increased risk for developing embryonic tumors. GPC3 is overexpressed in various cancers including hepatocellular carcinoma and melanoma. GPC3 is implicated in regulating many signaling pathways such as IGF, FGF, BMP and Wnt. GPC3 directly binds FGF­basic by the heparan sulfate chains. GPC3 has been considered as a tumor marker and a potential target for immunotherapy. 

 

 

Gene Symbol: GPC3; GPC-3; OCI5; SGB; DGSX; MXR7; SDYS; SGBS; OCI-5; SGBS1; GTR2-2

 

NCBI Gene ID: 2719

 

Uniprot Entry: P51654

 

Construct Details: Full length human GPC3 gene (encoding UniProt #P51654-1) is subcloned into the Lentiviral expression vector pLTC with an upstream CMV promoter and with or without 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. Multiple choices of a stable antibiotic selection marker are available.

 

Vector Type: pLTC or pLTC-IRES-Marker (lentiviral expression vector containing a heterologous CMV promoter with/without a selection 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: 1743 (bp)

 

Gene Insert Sequence:  

ATGGCCGGGACCGTGCGCACCGCGTGCTTGGTGGTGGCGATGCTGCTCAGCTTGGACTTCCCGGGACAGGCGCAGCCCCC

GCCGCCGCCGCCGGACGCCACCTGTCACCAAGTCCGCTCCTTCTTCCAGAGACTGCAGCCCGGACTCAAGTGGGTGCCAG

AAACTCCCGTGCCAGGATCAGATTTGCAAGTATGTCTCCCTAAGGGCCCAACATGCTGCTCAAGAAAGATGGAAGAAAAA

TACCAACTAACAGCACGATTGAACATGGAACAGCTGCTTCAGTCTGCAAGTATGGAGCTCAAGTTCTTAATTATTCAGAA

TGCTGCGGTTTTCCAAGAGGCCTTTGAAATTGTTGTTCGCCATGCCAAGAACTACACCAATGCCATGTTCAAGAACAACT

ACCCAAGCCTGACTCCACAAGCTTTTGAGTTTGTGGGTGAATTTTTCACAGATGTGTCTCTCTACATCTTGGGTTCTGAC

ATCAATGTAGATGACATGGTCAATGAATTGTTTGACAGCCTGTTTCCAGTCATCTATACCCAGCTAATGAACCCAGGCCT

GCCTGATTCAGCCTTGGACATCAATGAGTGCCTCCGAGGAGCAAGACGTGACCTGAAAGTATTTGGGAATTTCCCCAAGC

TTATTATGACCCAGGTTTCCAAGTCACTGCAAGTCACTAGGATCTTCCTTCAGGCTCTGAATCTTGGAATTGAAGTGATC

AACACAACTGATCACCTGAAGTTCAGTAAGGACTGTGGCCGAATGCTCACCAGAATGTGGTACTGCTCTTACTGCCAGGG

ACTGATGATGGTTAAACCCTGTGGCGGTTACTGCAATGTGGTCATGCAAGGCTGTATGGCAGGTGTGGTGGAGATTGACA

AGTACTGGAGAGAATACATTCTGTCCCTTGAAGAACTTGTGAATGGCATGTACAGAATCTATGACATGGAGAACGTACTG

CTTGGTCTCTTTTCAACAATCCATGATTCTATCCAGTATGTCCAGAAGAATGCAGGAAAGCTGACCACCACTATTGGCAA

GTTATGTGCCCATTCTCAACAACGCCAATATAGATCTGCTTATTATCCTGAAGATCTCTTTATTGACAAGAAAGTATTAA

AAGTTGCTCATGTAGAACATGAAGAAACCTTATCCAGCCGAAGAAGGGAACTAATTCAGAAGTTGAAGTCTTTCATCAGC

TTCTATAGTGCTTTGCCTGGCTACATCTGCAGCCATAGCCCTGTGGCGGAAAACGACACCCTTTGCTGGAATGGACAAGA

ACTCGTGGAGAGATACAGCCAAAAGGCAGCAAGGAATGGAATGAAAAACCAGTTCAATCTCCATGAGCTGAAAATGAAGG

GCCCTGAGCCAGTGGTCAGTCAAATTATTGACAAACTGAAGCACATTAACCAGCTCCTGAGAACCATGTCTATGCCCAAA

GGTAGAGTTCTGGATAAAAACCTGGATGAGGAAGGGTTTGAAAGTGGAGACTGCGGTGATGATGAAGATGAGTGCATTGG

AGGCTCTGGTGATGGAATGATAAAAGTGAAGAATCAGCTCCGCTTCCTTGCAGAACTGGCCTATGATCTGGATGTGGATG

ATGCGCCTGGAAACAGTCAGCAGGCAACTCCGAAGGACAACGAGATAAGCACCTTTCACAACCTCGGGAACGTTCATTCC

CCGCTGAAGCTTCTCACCAGCATGGCCATCTCGGTGGTGTGCTTCTTCTTCCTGGTGCACTGA

 

 

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. Nat. Genet. 12: 241 (1996)

2. J. Biol. Chem. 272:7574 (1997)

3. J. Clinical Invest. 108:497 (2001) 

4. J. Cell. Biol. 163: 625 (2003)

5. Clin Cancer Res. 10: 6612 (2004)

 

 

 

 

Product datasheet (pdf) can be downloaded here: LTP2058-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.