TPST2-EGFP
(Plasmid #66618)

• Purpose: Encodes GFP fusion of TPST2 trans Golgi marker (Tyrosyl protein sulfotransferase 2)
• Depositing Lab: David Stephens
• Publication: Spooner et al Biochem J. 2008 Sep 15;414(3):471-84. doi: 10.1042/BJ20080149.

Backbone

• Vector backbone: pEGFP-N1
• Backbone manufacturer: Clontech
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: TPST2
• Alt name: Homo sapiens tyrosylprotein sulfotransferase 1
• Species: H. sapiens (human)
• Mutation: Insert contains T370A relative to NM_001008566.1
• GenBank ID: NM_001008566.1
• Entrez Gene: TPST2 (a.k.a. TANGO13B, TPST-2)

Resource Information

• A portion of this plasmid was derived from a plasmid made by: RZPD, now distributed through Source Bioscience. Was obtained direct from RZPD in our case
• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Molecular cloning of TPST (tyrosylprotein sulfotransferase)–EGFP (enhanced green fluorescent protein)
The TPST1 and TPST2 sequences were obtained from human ESTs (expressed sequence tags) from the RZPD (Deutsches Ressourcenzentrum für Genomforschung, Berlin, Germany; GenBank® accession numbers NM_003596 and AF061254 respectively). TPST1–EGFP and TPST2–EGFP were obtained by PCR using the following primers: plus strand, 5′-GGGGCTCGAGATGGTTGGAAAGCTGAAG-3′, and minus strand, 5′-GGGGCTGCAGCTCCACTTGCTCAGTC-3′ for TPST1; and plus strand, 5′-GGGGCTCGAGATGCGCCTGTCGGTGC-3′, and minus strand, 5′-GGGGCTGCAGCGAGCTTCCTAAGTGG-3′ for TPST2 (XhoI and PstI, restriction sites are underlined). PCR products were purified and ligated into the pGEM-T-Easy vector (Promega) following the manufacturer's instructions. Positive clones were digested with XhoI and PstI, and the fragments obtained were inserted in the corresponding sites of pEGFP-N1 (Clontech). All constructs were verified by double-strand DNA sequencing (MWG Biotech, Ebersburg, Germany). Two differences to the published sequence of TPST2 were identified (276C>T and 1002T>C, relative to the start codon), but neither of which affect the predicted protein sequence.

Insert contains T370A relative to reference sequence. The depositing laboratory does not know the effect of this substitution on gene function