Research Tools

Gene Constructs and Promoters

The following gene constructs and promoters are available from the lab of Jeffrey A. Whitsett, MD:

  • ~3.7 Kb human SP-C (w/maps and sequence)
  • ~2.0 Kb mouse CCSP (w/maps and sequence)
  • ~4.8 Kb mouse SP-C (w/maps and sequence)
  • ~2.2 Kb rat CCSP (w/maps and sequence)
  • 3.7 hSP-C-rtTA-SV40-t-intron poly A
  • 2.2 rCCSP-rtTA-human growth hormone poly A 3'UT
  • (otet) 7 CMV-bovine growth hormone poly A 3'UT
  • (otet) 7 CMV-SV40-t-intron poly A
  • SP-C promoter
  • Mouse 4.8 kb SP-C promoter
  • Human 3.7 kb SP-C promoter
  • Mouse CCSP promoter
  • Rat CCSP promoter
  • CC10 Promoter and chitinase + MMP-7
  • SP-B-luciferase and TTF-1 luciferase constructs
  • TTF-1 promoter
  • SP-C DNA construct

The following gene constructs and promoters are available from the lab of Jeffrey Robbins, PhD:

  • Transcriptional apparatus for driving transgenic expression in the heart, including:
    • the alpha myosin heavy chain promoters (mouse and rat)
    • The beta myosin heavy chain promoter (mouse and rabbit)
    • The myosin light chain promoter for ventricle specific expression (mouse)
    • The minimal tet-responsive activator (basic on Gossen and Bujard, PNAS, 1992)
    • The responder line, developed through a grant from P&G. (Generally useful for inducible expression in transgenic animals)

The following gene constructs and promoters are available from the lab of Cong Yan, PhD:

  • Rat TTF-1-FLAG wild type (U1-D1) expression vector

The following gene constructs and promoters are available from the lab of David Repaske, PhD, MD:

  • PDE1B constructs

The following promoter is available from the lab of Kathleen P. Anderson, PhD:

  • LMO2 promoter (drives hematopoetic-specific expression)

The following adenoviral vectors are available from the lab of Jeffery Molkentin, PhD:

#All were plaque purified and expanded using standard techniques.
#All were Western blot tested for protein expression in cultured neonatal cardiomyocytes or C2C12 cells.
#Many of the dominant negatives were also tested for inhibitory activity.
#Many of the adenoviral vectors from the Molkentin Lab are now available at Seven Hills Bioreagents, a venture of Cincinnati Children's Research Foundation.

  • activated calcineurin (amino acids 1-398) Mouse cDNA for truncated CnA-beta (Western tested)
  • activated calcineurin Flag (amino acids 1-398) Mouse cDNA for truncated CnA-beta with 5' Flag epitope (Western tested)
  • calcineurin inhibitory cain peptide (amino acids 1989-2182) Mouse cDNA fragment with 5' flag epitope (Western tested)
  • calcineurin inhibitory AKAP79 peptide (amino acids 60-358) Human cDNA fragment with 5' flag epitope (Western tested)
  • bcl-XL (wild type) Human cDNA (Western tested)
  • consititutively nuclear NFATc4 (amino acids 317-902) Human cDNA fragment (Western tested)
  • consistutively nuclear NFATc3 (amino acids 282-1068) Human cDNA fragment with 5' flag tag (Western tested)
  • GATA4 wild type Mouse cDNA (Western tested)
  • GATA4 anti-sense (potential dominant negative) Mouse cDNA in reverse (Western tested to reduce endogenous GATA4)
  • GATA4-engrailed dominant negative fusion (GATA4 211-329 - engr 2-298) Mouse GATA4 cDNA fused to amino acids 2-298 of engrailed cDNA with 5' flag epitope. (Western tested)
  • MEK1 activated (Ser 217/221 Glu) Rabbit cDNA, (Western tested to induce ERK1/ 2 phos)
  • MEK1 dominant negative (Ser 221 Ala) Rabbit cDNA, (Western tested to block ERK1/ 2 phos)
  • MKK6 activated (Ser 207/211 Glu) Human cDNA (Western tested to induce p38 phos)
  • p38alpha wild type Human cDNA with 5'Flag epitope (Western tested)
  • p38beta wild type Human cDNA with 5'Flag epitope (Western tested)
  • p38alpha dominant negative (AGF mutation) Human cDNA with 5'Flag epitope(Western tested to block endogenous p38 phos)
  • p38beta dominant negative (AGF mutation) Human cDNA with 5'Flag epitope(Western tested to block endogenous p38 phos)
  • mkk3 dominant negative (Ser 189/193 Ala) Human cDNA with 5'Flag epitope (Western tested to block p38 phos)
  • p38alpha-GFP fusion protein (cell localization studies) Human p38alpha cDNA fused to enhanced GFP(Immunflor. Tested)
  • p38beta-GFP fusion protein (cell localization studies) Human p38beta cDNA fused to enhanced GFP (Immunflor. Tested)
  • p38alpha-mutant estrogen receptor fusion (fusion protein activated by tamoxifen) Human p38alpha cDNA fused to mutant estrogen receptor (Western tested)
  • p38beta-mutant estrogen receptor fusion (fusion protein activated by tamoxifen) Human p38beta cDNA fused to mutant estrogen receptor (Western tested)
  • JNK1 wild type Mouse cDNA (Western tested)
  • JNK2 wild type Mouse cDNA (Western tested)
  • JNK1 dominant negative (APF mutation) Human cDNA with 5'Flag epitope (Western tested)
  • JNK2 dominant negative (APF mutation) Human cDNA with 5' HA epitope (Western tested)
  • MKK4 dominant negative (Lys 129 Arg) Human cDNA (Western tested)
  • MKP-1 phosphatase Mouse cDNA (Western tested to dephosphorylate p38 and JNK)
  • M3/6 phosphatase (specific for JNK dephosphorylation) Mouse cDNA (Western tested to dephosphorylate JNK perferentially)
  • TAM67 c-Jun dominant negative (blocks c-Jun transcriptional activity Human cDNA (Western blot tested)
  • ERK2 wild type (5' Flag tagged) Rat cDNA, but same amino acids as Human and mouse (Western tested)
  • ERK2 dominant negative. (AEF mutation) (5' Flag Tagged) Rat cDNA, for dominant negative extracellular signal-regulated kinase cDNA with 5' flag tag
  • ERalpha wildtype (estrogen receptor) Human cDNA (Western tested)
  • ERbeta (wildtype beta estrogen receptor, human cDNA) (Western tested)
  • N17Ras (dominant negative) Human cDNA (Western tested)
  • PKC7-alpha (activated truncation of PKC alpha) rabbit cDNA (Western tested)
  • AdLacZ-nls (beta-galactosidase, nuclear localized) Bacterial protein product (tested by staining of infected cells)
  • PKCalpha (wildtype rabbit PKCalpha cDNA) (Western tested)
  • PKCalpha-dn (dominant negative PKCalpha mutant cDNA) (Western tested)
  • PKCbetaII (wildtype mouse PKCbetaII cDNA) (Western tested)
  • PKCbetaII-dn (dominant negative PKCbetaII mutant cDNA) (Western tested)
  • PKCepsilon (wildtype mouse PKCepsilon cDNA) (Western tested)
  • PKCepsilon-dn (dominant negative PKCepsilon mutant cDNA) (Western tested)
  • PKCdelta (wildtype mouse PKCdelta cDNA) (Western tested)
  • PKCdelta-dn (dominant negative PKCdelta mutant cDNA) (Western tested)
  • PKCzeta (wildtype mouse PKCzeta cDNA) (Western tested)
  • PKCzeta-dn (dominant negative PKCzeta mutant cDNA) (Western tested)
  • PDK-1 (wildtype PDK-1 cDNA) (Western tested)
  • MEF2A (wildtype MEF2A transcription factor, Human cDNA) (Western tested)
  • MEF2B (wildtype MEF2B transcription factor, mouse cDNA) (Western tested)
  • MEF2C (wildtype MEF2C transcription factor, mouse cDNA) (Western tested)
  • MEF2D (wildtype MEF2D transcription factor, mouse cDNA) (Western tested)
  • MEF2C-DN (Dominant negative mutant of MEF2C, mouse cDNA) (Western tested)
  • CnB (calcineurin B regulatory subunit, mouse cDNA) (Western tested)
  • MKP-3 (MAPK phosphatase –3, ERK1/2 specific) (mouse cDNA) (Western tested and shown to reverse ERK1/2 activity)
  • NFAT-luciferase reporter (reporter adenovirus for NFAT activity and calcineurin activation)
  • NFATc3 (full-length NFATc3, human cDNA) (Western tested)
  • dHAND (transcription factor, mouse cDNA)
  • Nkx2.5 (transcription factor, mouse cDNA) (Western tested)
  • HDAC4 (histone deacetylase 4) (human cDNA)