Plant Plasma Membrane Proton Pump Immunocytochemical Visualization W/ Tem

Abstract

The plasma membrane proton pump (H+-ATPase) in plants and fungi converts chemical energy into electrical energy, and generates the protonmotive force used to transport solutes into and out of all cells. My laboratory has cloned twelve different genes encoding the ATPase in A. thaliana, and is using light and electron microscopy techniques to identify their tissue and intracellular localization. When standard immunofluoresces microscopy is not sufficient for immunolocalization of antigens we will use the confocal microscope at the IMR. We will high pressure freeze samples for better preservation of antigenicity. After freezing the samples will be freeze substituted, embedded, sectioned and immunogold labeled. Immunolabeling will be observed by TEM. For this purpose, we are using two antibody preparations ( I ) a standard approach, using antibody generated against proteins obtained via expression in E. coli, (2) generation of recombinant plants containing a foreign 'epitope' (either mammalian c-myc or bacterial glucuronidase) attached to the carboxy terminus of the ATPase polypeptide. Our present studies focus on an ATPase gene isoform which is expressed in phloem cells.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Center for Research Resources (NCRR)
• Type: Biotechnology Resource Grants (P41)
• Project #: 3P41RR000570-27S1
• Application #: 2757536
• Budget Start: 1997-10-01
• Budget End: 1998-09-30
• Support Year: 27
• Fiscal Year: 1998

Institution

• Name: University of Wisconsin Madison
• DUNS #: 161202122
• City: Madison
• State: WI
• Country: United States
• Zip Code: 53715

Publications

Malecki, Marek; Putzer, Emily; Sabo, Chelsea et al. (2014) Directed cardiomyogenesis of autologous human induced pluripotent stem cells recruited to infarcted myocardium with bioengineered antibodies. Mol Cell Ther 2:

Malecki, Marek (2014) 'Above all, do no harm': safeguarding pluripotent stem cell therapy against iatrogenic tumorigenesis. Stem Cell Res Ther 5:73

Mavroudi, Maria; Zarogoulidis, Paul; Porpodis, Konstantinos et al. (2014) Stem cells' guided gene therapy of cancer: New frontier in personalized and targeted therapy. J Cancer Res Ther (Manch) 2:22-33

Malecki, Marek; LaVanne, Christine; Alhambra, Dominique et al. (2013) Safeguarding Stem Cell-Based Regenerative Therapy against Iatrogenic Cancerogenesis: Transgenic Expression of DNASE1, DNASE1L3, DNASE2, DFFB Controlled By POLA1 Promoter in Proliferating and Directed Differentiation Resisting Human Autologous Pluripotent J Stem Cell Res Ther Suppl 9:

Malecki, Marek; Tombokan, Xenia; Anderson, Mark et al. (2013) TRA-1-60(+), SSEA-4(+), POU5F1(+), SOX2(+), NANOG(+) Clones of Pluripotent Stem Cells in the Embryonal Carcinomas of the Testes. J Stem Cell Res Ther 3:

Malecki, Marek (2013) Improved targeting and enhanced retention of the human, autologous, fibroblast-derived, induced, pluripotent stem cells to the sarcomeres of the infarcted myocardium with the aid of the bioengineered, heterospecific, tetravalent antibodies. J Stem Cell Res Ther 3:

Malecki, Marek; Dahlke, Jessica; Haig, Melissa et al. (2013) Eradication of Human Ovarian Cancer Cells by Transgenic Expression of Recombinant DNASE1, DNASE1L3, DNASE2, and DFFB Controlled by EGFR Promoter: Novel Strategy for Targeted Therapy of Cancer. J Genet Syndr Gene Ther 4:152

Zarogoulidis, Paul; Darwiche, Kaid; Sakkas, Antonios et al. (2013) Suicide Gene Therapy for Cancer - Current Strategies. J Genet Syndr Gene Ther 4:

Malecki, Marek; Sabo, Chelsea; Putzer, Emily et al. (2013) Recruitment and retention of human autologous CD34+ CD117+ CD133+ bone marrow stem cells to infarcted myocardium followed by directed vasculogenesis: Novel strategy for cardiac regeneration. Mol Cell Ther 1:

Malecki, Marek; Malecki, Bianca (2012) Routing of Biomolecules and Transgenes' Vectors in Nuclei of Oocytes. J Fertili In Vitro 2012:108-118