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Marker Gene Monthly Newsletter

October, 2002

Volume 2, Number 10

© Copyright MGT, Inc., 2007. Published by Marker Gene Technologies, Inc., The University of Oregon Riverfront Research Park, 1850 Millrace Drive, Eugene, Oregon 97403-1992 USA. All rights reserved. For information on the use or copying of the material contained in this document, please contact us at techservice@markergene.com. Please see below for subscription information and updates. This newsletter is labeled as an ADVERTISEMENT in accordance with the CAN-SPAM act of 2003, S.877 Public Law: 108-187.

"Jet-Lag" and Circadian Rhythms in Plants.

The effects of daylight length and time-changes, and the physiological events that occur in eukaryotic cells in response to these events, have long intrigued scientists. Some of these processes are beginning to be understood in plants and animals by cloning marker genes (for example firefly luciferase, luc) into cells under control of specific circadian promoter elements. By adding the reagent D-luciferin (M0237) to these cloned tissues, Steven Kay’s lab at the Scripps Research Institute in San Diego (see: http://www.scripps.edu/cb/kay/) has identified several new genes that “light up”, i.e. are responsible for the physiological changes in plants and animals, when daylight and night-time cycled gene expression occurs. For more information, see the references below:

Covington, M.F., Panda, S., Liu, X.L., Strayer, C.A., Kay, S.A., Wagner, D.R. (2001) ELF3 modulates resetting of the circadian clock in Arabidopsis. The Plant Cell 13, 1305-1316.

Millar A.J. and Kay S.A. (1996). Integration of circadian and phototransduction pathways in the network controlling CAB gene transcription in Arabidopsis. Proc. Natl. Acad. Sci. USA, 93, 15491-15496.

Millar A.J., Straume M., Chory J., Chua N.-H. and Kay S.A. (1995) The regulation of circadian period by phototransduction pathways in Arabidopsis. Science, 267, 1163-1166.

FDG Staining of live Zebrafish Embryos
Over the last several years, the zebrafish has emerged as an attractive model for vertebrate developmental biology, largely because of the ease of genetic analysis and the transparent nature of the embryo. However, despite many advantages, zebrafish researchers still require certain experimental tools, including viable embryonic markers for genetic and cell lineage studies. Staining with the fluorescent b-galactosidase substrate FDG (M0250) is a popular method of detecting transgenic activity in these embryos when using the lacZ reporter gene, but may require dechorionation of the embryos prior to staining. See the references below for more information about these techniques.

Rossant, J., Hopkins, N. (1992) Of fin and fur: mutational analysis of vertebrate embryonic development.

Genes Devel. 6:1–13.Lin S, Yang S, Hopkins N. lacZ expression in germline transgenic zebrafish can be detected in living embryos. Dev. Biol. (1994) 161(1):77-83

Oxidative Burst in Neutrophils with DHR123

The reduced or missing oxidative burst activity in leukocytes is an indication of hereditary diseases like chronic granulomatous disease (CGD). Treatment of cells (e.g. heparinized whole blood) with the reduced dye DHR123, (dihydrorhodamine 123, M0545) is a sensitive assay for oxidative activity in such cells, with allied induction using either the chemotactic peptide N-formyl-Met-Leu-Phe (fMLP), the protein kinase C ligand phorbol 12-myristate-13-acetate (PMA) or bacterial challenge. DHR123 is practically non-fluorescent until oxidized intracellularly to the bright red fluorescent rhodamine 123 product. For more information see the references listed below or visit our Website:

Rothe G, Oser A & G. Valet. (1988) Dihydrorhodamin 123: a new flow cytometric indicator for respiratory burst activity in neutrophil granulocytes. Naturwissenschaften 75: 354 - 355.

Dobmeyer, T.S., Raffel, B. Dobmeyer, J.M., Findhammer, S., Klein, S.A., Kabelitz, D. Hoelzer, D., Helm, E.B. & Rossol.(1995) Decreased function of monocytes and granulocytes during HIV-1 infection correlates with CD4 cell counts. Eur. J. Med. Res. 1: 9-15.

Gessler, P., Nebe, T. Birle, A., Haas, N. & W. Kachel. (1996) Neutrophil respiratory burst in term and preterm neonates without signs of infection and in those with increased levels of C-Reactive Protein. Pediatr. Res. 39: 843-848.

Elbim, C., Chollet-Martin, S., Bailly, S., Hakim, J. & M.A. Gougerot-Pocidalo. (1993) Priming of polymorphonuclear neutrophils by tumor necrosis factor in whole blood: Identification of two polymorphonuclear neutrophil subpopulations in response to formyl-peptides. Blood 82: 663-640.

Adoptive Immunotherapy for Melanomas
Steven Rosenberg’s laboratory at the National Cancer Institute recently reported results of a new study were reactive “killer” T-cells were removed from patients’ tumor regions, and grown in culture to high density. When re-infused into these same patients, their ability to mount an immune response to the tumors was highly improved. The key factors of cell viability (and proliferation) during culturing and after re-infusion were addressed using a regimen where the patient’s own immune system was depleted before transfer (prior lymphodepletion using a combination of drugs). These techniques present a potential promising new weapon for the treatment of cancer using the body’s own immune system.

Dudley et al. (2002) Cancer Regression and Autoimmunity in Patients After Clonal Repopulation with Antitumor Lymphocytes. Science 298 (5594): 850-854.

Winter, H., Fox, B.A., Adoptive Cellular Immunotherapy of Cancer. Current Opinion in Molecular Therapeutics. 1 (1):89-97,1999.

Black Spot Disease in Citrus Fruits

Black Spot is a fungal disease caused by Guignardia citricarpa. It causes black lesions on citrus fruits like oranges and grapefruits, and is becoming a threat to the US agriculture industry. The disease has not been reported in U.S. citrus producing states, but has been found in parts of Australia, South Africa and Argentina and is becoming a serious problem in Brazil. Importation or export of fruits from the US are in jeopardy since the European Union (EU) and South Africa in 2000 now threaten to slow citrus exports because of zealous application of protective standards. Recently several assays including a definitive PCR-based assay have been developed, which may alleviate these quarantine issues. For more information on the disease and these assays, see the references below:

Baayen, R.P. , (2002) “Nopathogenic isolates of the citrus black spot fungus, Guignardia citricarpa, identified as a cosmopolitan endophyte of woody plants, G. mangiferae (Phyllosticta capitalensis)”, Phytopathology 92 (5): 464-477.

New Catalog Will Be Available Soon

The 2003-2004 edition of the Marker Gene catalog is heading to the printers. Many new products and kits, additional literature references, data and protocols will be included, as well as new information about our old products. Be sure to add your name to our mailing list. Please visit our Web site and fill out our Customer Information Form, or e-mail us at techservice@markergene.com and we will have a copy sent out to you.
Sign up now!

Compare Our Quality.
Marker Gene strives to offer our customers products of the highest quality and at the best possible prices. Our years of experience allow us to provide timely products for less cost to you. See our latest Price Comparison Chart that compares our prices with those from several alternate sources, to see if you can save money by switching to Marker Gene (http://www.markergene.com/crossref.htm). Or visit our website at www.markergene.com and click on the link “COMPARE”. We think you will appreciate our efforts to keep costs low and maintain excellent quality of our products for your research. For more information about any of our products, simply telephone us toll free at 1-888-218-4062 or contact us by e-mail at techservice@markergene.com. We will be happy to send you more about our products and their specifications.

CONTRACT RESEARCH@markergene.com

Marker Gene Technologies, Inc. has the expertise to perform contract research with you on your project. We have worked with many biotechnology and pharmaceutical companies on successful, proprietary and patented projects.

Contract Research and Development Capabilities in the following areas:

Established in 1993 at the University of Oregon Riverfront Research Park.

Screening Assay Development for HTS and uHTS

Chemical and Cellular Assays – High-Content Screening.

DNA/RNA (genomics) and protein (proteomics) labeling and assay development.

Pharmaceutical Intermediates - design, synthesis, and in vitro testing in mammalian cell culture.

Specializing in Carbohydrate, Lipid, Peptide, and Nucleic Acid Chemistries.

Fully equipped laboratories (Biochemistry, Chemical Synthesis, Tissue Culture, Analytical).

Confidentiality, help in patent preparation and filings.

Contact us by telephone at (888) 218-4062 or (541) 342-3760 or FAX us at (541) 342-1960 or you can write to us at Contract Research, Marker Gene Technologies, Inc., 1850 Millrace Drive, Eugene, Oregon 97403-1992 or contact us by e-mail at: techservice@markergene.com

Marker Gene Accepts Major Credit Cards.

Place your orders now, using Master Card or Visa and save time and money! Our Customer Assistance Staff can now accept either Master Card or Visa Credit Card orders, securely by telephone (toll-free) at 1-888-218-4062 (Domestic orders only). We will continue to accept Institutional Purchase Orders for our products, online or by FAX at 1-541-342-1960. International customers should contact us by e-mail, post or telephone for more information about International Distributors and ordering. For information on pricing for individual products, or for a quote on bulk quantities of our products or kits, please contact our technical assistance staff at techservice@markergene.com. We will be happy to assist you.

To add your name to our WebNewsletter mailing list, please use the following link: subscribe@markergene.com.

To unsubscribe from our mailing list, please use the following link: unsubscribe@markergene.com. Your e-mail address will be deleted in 2-3 business days.


	If you are having trouble viewing this email, click on this link. To order any of our products, please go to our ORDER FORM or visit one of our distributors:
	Marker Gene Monthly Newsletter October, 2002 Volume 2, Number 10 © Copyright MGT, Inc., 2007. Published by Marker Gene Technologies, Inc., The University of Oregon Riverfront Research Park, 1850 Millrace Drive, Eugene, Oregon 97403-1992 USA. All rights reserved. For information on the use or copying of the material contained in this document, please contact us at techservice@markergene.com. Please see below for subscription information and updates. This newsletter is labeled as an ADVERTISEMENT in accordance with the CAN-SPAM act of 2003, S.877 Public Law: 108-187.
	"Jet-Lag" and Circadian Rhythms in Plants. The effects of daylight length and time-changes, and the physiological events that occur in eukaryotic cells in response to these events, have long intrigued scientists. Some of these processes are beginning to be understood in plants and animals by cloning marker genes (for example firefly luciferase, luc) into cells under control of specific circadian promoter elements. By adding the reagent D-luciferin (M0237) to these cloned tissues, Steven Kay’s lab at the Scripps Research Institute in San Diego (see: http://www.scripps.edu/cb/kay/) has identified several new genes that “light up”, i.e. are responsible for the physiological changes in plants and animals, when daylight and night-time cycled gene expression occurs. For more information, see the references below: Covington, M.F., Panda, S., Liu, X.L., Strayer, C.A., Kay, S.A., Wagner, D.R. (2001) ELF3 modulates resetting of the circadian clock in Arabidopsis. The Plant Cell 13, 1305-1316. Millar A.J. and Kay S.A. (1996). Integration of circadian and phototransduction pathways in the network controlling CAB gene transcription in Arabidopsis. Proc. Natl. Acad. Sci. USA, 93, 15491-15496. Millar A.J., Straume M., Chory J., Chua N.-H. and Kay S.A. (1995) The regulation of circadian period by phototransduction pathways in Arabidopsis. Science, 267, 1163-1166.
	FDG Staining of live Zebrafish Embryos Over the last several years, the zebrafish has emerged as an attractive model for vertebrate developmental biology, largely because of the ease of genetic analysis and the transparent nature of the embryo. However, despite many advantages, zebrafish researchers still require certain experimental tools, including viable embryonic markers for genetic and cell lineage studies. Staining with the fluorescent b-galactosidase substrate FDG (M0250) is a popular method of detecting transgenic activity in these embryos when using the lacZ reporter gene, but may require dechorionation of the embryos prior to staining. See the references below for more information about these techniques. Rossant, J., Hopkins, N. (1992) Of fin and fur: mutational analysis of vertebrate embryonic development. Genes Devel. 6:1–13.Lin S, Yang S, Hopkins N. lacZ expression in germline transgenic zebrafish can be detected in living embryos. Dev. Biol. (1994) 161(1):77-83
	Oxidative Burst in Neutrophils with DHR123 The reduced or missing oxidative burst activity in leukocytes is an indication of hereditary diseases like chronic granulomatous disease (CGD). Treatment of cells (e.g. heparinized whole blood) with the reduced dye DHR123, (dihydrorhodamine 123, M0545) is a sensitive assay for oxidative activity in such cells, with allied induction using either the chemotactic peptide N-formyl-Met-Leu-Phe (fMLP), the protein kinase C ligand phorbol 12-myristate-13-acetate (PMA) or bacterial challenge. DHR123 is practically non-fluorescent until oxidized intracellularly to the bright red fluorescent rhodamine 123 product. For more information see the references listed below or visit our Website: Rothe G, Oser A & G. Valet. (1988) Dihydrorhodamin 123: a new flow cytometric indicator for respiratory burst activity in neutrophil granulocytes. Naturwissenschaften 75: 354 - 355. Dobmeyer, T.S., Raffel, B. Dobmeyer, J.M., Findhammer, S., Klein, S.A., Kabelitz, D. Hoelzer, D., Helm, E.B. & Rossol.(1995) Decreased function of monocytes and granulocytes during HIV-1 infection correlates with CD4 cell counts. Eur. J. Med. Res. 1: 9-15. Gessler, P., Nebe, T. Birle, A., Haas, N. & W. Kachel. (1996) Neutrophil respiratory burst in term and preterm neonates without signs of infection and in those with increased levels of C-Reactive Protein. Pediatr. Res. 39: 843-848. Elbim, C., Chollet-Martin, S., Bailly, S., Hakim, J. & M.A. Gougerot-Pocidalo. (1993) Priming of polymorphonuclear neutrophils by tumor necrosis factor in whole blood: Identification of two polymorphonuclear neutrophil subpopulations in response to formyl-peptides. Blood 82: 663-640.
	Adoptive Immunotherapy for Melanomas Steven Rosenberg’s laboratory at the National Cancer Institute recently reported results of a new study were reactive “killer” T-cells were removed from patients’ tumor regions, and grown in culture to high density. When re-infused into these same patients, their ability to mount an immune response to the tumors was highly improved. The key factors of cell viability (and proliferation) during culturing and after re-infusion were addressed using a regimen where the patient’s own immune system was depleted before transfer (prior lymphodepletion using a combination of drugs). These techniques present a potential promising new weapon for the treatment of cancer using the body’s own immune system. Dudley et al. (2002) Cancer Regression and Autoimmunity in Patients After Clonal Repopulation with Antitumor Lymphocytes. Science 298 (5594): 850-854. Winter, H., Fox, B.A., Adoptive Cellular Immunotherapy of Cancer. Current Opinion in Molecular Therapeutics. 1 (1):89-97,1999.
	Black Spot Disease in Citrus Fruits Black Spot is a fungal disease caused by Guignardia citricarpa. It causes black lesions on citrus fruits like oranges and grapefruits, and is becoming a threat to the US agriculture industry. The disease has not been reported in U.S. citrus producing states, but has been found in parts of Australia, South Africa and Argentina and is becoming a serious problem in Brazil. Importation or export of fruits from the US are in jeopardy since the European Union (EU) and South Africa in 2000 now threaten to slow citrus exports because of zealous application of protective standards. Recently several assays including a definitive PCR-based assay have been developed, which may alleviate these quarantine issues. For more information on the disease and these assays, see the references below: Baayen, R.P. , (2002) “Nopathogenic isolates of the citrus black spot fungus, Guignardia citricarpa, identified as a cosmopolitan endophyte of woody plants, G. mangiferae (Phyllosticta capitalensis)”, Phytopathology 92 (5): 464-477.
	New Catalog Will Be Available Soon The 2003-2004 edition of the Marker Gene catalog is heading to the printers. Many new products and kits, additional literature references, data and protocols will be included, as well as new information about our old products. Be sure to add your name to our mailing list. Please visit our Web site and fill out our Customer Information Form, or e-mail us at techservice@markergene.com and we will have a copy sent out to you. Sign up now!
	Compare Our Quality. Marker Gene strives to offer our customers products of the highest quality and at the best possible prices. Our years of experience allow us to provide timely products for less cost to you. See our latest Price Comparison Chart that compares our prices with those from several alternate sources, to see if you can save money by switching to Marker Gene (http://www.markergene.com/crossref.htm). Or visit our website at www.markergene.com and click on the link “COMPARE”. We think you will appreciate our efforts to keep costs low and maintain excellent quality of our products for your research. For more information about any of our products, simply telephone us toll free at 1-888-218-4062 or contact us by e-mail at techservice@markergene.com. We will be happy to send you more about our products and their specifications.
	CONTRACT RESEARCH@markergene.com Marker Gene Technologies, Inc. has the expertise to perform contract research with you on your project. We have worked with many biotechnology and pharmaceutical companies on successful, proprietary and patented projects. Contract Research and Development Capabilities in the following areas: Established in 1993 at the University of Oregon Riverfront Research Park. Screening Assay Development for HTS and uHTS Chemical and Cellular Assays – High-Content Screening. DNA/RNA (genomics) and protein (proteomics) labeling and assay development. Pharmaceutical Intermediates - design, synthesis, and in vitro testing in mammalian cell culture. Specializing in Carbohydrate, Lipid, Peptide, and Nucleic Acid Chemistries. Fully equipped laboratories (Biochemistry, Chemical Synthesis, Tissue Culture, Analytical). Confidentiality, help in patent preparation and filings. Contact us by telephone at (888) 218-4062 or (541) 342-3760 or FAX us at (541) 342-1960 or you can write to us at Contract Research, Marker Gene Technologies, Inc., 1850 Millrace Drive, Eugene, Oregon 97403-1992 or contact us by e-mail at: techservice@markergene.com
	Marker Gene Accepts Major Credit Cards. Place your orders now, using Master Card or Visa and save time and money! Our Customer Assistance Staff can now accept either Master Card or Visa Credit Card orders, securely by telephone (toll-free) at 1-888-218-4062 (Domestic orders only). We will continue to accept Institutional Purchase Orders for our products, online or by FAX at 1-541-342-1960. International customers should contact us by e-mail, post or telephone for more information about International Distributors and ordering. For information on pricing for individual products, or for a quote on bulk quantities of our products or kits, please contact our technical assistance staff at techservice@markergene.com. We will be happy to assist you.
	To add your name to our WebNewsletter mailing list, please use the following link: subscribe@markergene.com. To unsubscribe from our mailing list, please use the following link: unsubscribe@markergene.com. Your e-mail address will be deleted in 2-3 business days.