Marker Gene Monthly Newsletter   

January, 2003

Volume 3, Number 1

© 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.

Higher Transfection Efficiency With Introns.

Recent work from Melissa Moore’s laboratory at the Howard Hughes Medical Institute and Brandeis University in Waltham, MA have indicated that exon splicing of intron sequences leave behind signature proteins on mRNA that improve expression levels in vivo over cDNA-prepared mRNA without introns.  The reasons for the improved mRNA expression are several, but may include better transport from the nucleus, lower RNA degradation rates as well as improved ribosome or ER trafficking.  For more information about improving expression levels using intron-added vector sequences, see the following refereneces:

• Le Hir H, Gatfield D, Izaurralde E and Moore MJ. (2001). “The exon-exon junction complex provides a binding platform for factors involved in mRNA export and NMD.” EMBO J 20:4987-97.
• Moore MJ. (2002) “RNA events. No end to nonsense.” Science 11;298(5592):370-1
  

Live Cell Firefly Luciferase Assays.

Analysis of cloned Luciferase (luc) activity in live mammalian cells can be accomplished using a variety of methods, including lysis assays and microtiterplate systems, but it is often useful to recover the cells under analysis for later expansion or secondary analysis.  Our Luciferase Assay Kit (M0626) contains the reagents, buffers and a detailed protocol for live cell luc analysis in culture, by microscopic analysis, microplate analysis or using the common lysis assay conditions.  The kit contains sufficient reagents for up to 1000 assays, using the conditions described.  For more information about these assays see the references below or visit our Web site.

• Maechler, P., Wang, H., Wollheim, C.B. “Continuous monitoring of ATP levels in living insulin secreting cells expressin cytosolic firefly luciferase.” (1998) FEBS Lett. 422: 328-332.
• Takasuka, N., White, M.R.H., Wood, C.D., Robertson, W.R., Davis, J.R.E., “Dynamic Changes in Prolactin Promoter Activation in Individual Living Lactotrophic Cells.” (1998) Endocrinology 139(3): 1361-1368.

New Proteomics Hydrophobicity Assay for SDS-PAGE Gels.

The isolation of hydrophobic membrane proteins can result in complex protein mixtures that are difficult to purify, identify, and quantitate by bioinformatics.  Utilizing 2D gel electrophoresis analysis, individual bands can typically be isolated but may contain multiple protein samples or a variety of functional components.  Our new Hydrophobic Protein Analysis Kit (M0794) uses 2-p-toluidinylnapthalene-6-sulfonic acid (TNS, M0704) as the protein-detecting reagent.  TNS is one of a class of compounds that fluoresce strongly when bound to specific native proteins, based upon their hydrophobic surface area.  The kit contains reagents and a detailed protocol for solution 1D or 2D SDS-PAGE analysis of hydrophobic proteins in complex samples.  For more information see the references below.

• McClure, W.O., and Edelman, G.M. (1966), Biochem. 5(6): 1908-1919.
• Neff, T.L., Naleway, J.J., “Development of a Sensitive Proteomic Hydrophobicity Assay Based on 2-p-Toluidinylnapthalene-6-Sulfonic Acid (TNS) as the Protein-Detecting Reagent.” (2003) Anal. Biochem. (submitted for publication).

New Plant Selection System Using Mannose.

A new selection marker that makes use of the inability of most plants to metabolize the simple sugar mannose, has been developed by Syngenta AG, Basel, Switzerland. Transgenic plants expressing the enzyme phosphomannose isomerase (PHI) encoded by the manA gene from E. coli are able to convert mannose-6-phosphate to fructose-6-phosphate, which can then be utilized. The technology has been shown to be effective in a range of commercially important crops and plant models.  For more information about the techniques and system, see the references below:

• Miles, J.S., Guest, J.R., “Nucleotide sequence and transcriptional start point of the phosphomannose isomerase gene (manA) of Escherichia coli.” (1984) Gene 32: 41-48.
• Joersbo, M., Donaldson, I., Kreiberg, J., Petersen, S.G., Brundstedt, J.,  Okkels, F.T., (1998) ”Analysis of mannose selection used for transformation of sugar beet” Molecular Breeding 4:111-117
•   Wang, A.S., Evans, R.A., Altendorf, P.R., Hanten, J.A., Doyle, M.C., Rosichan, J.L., (2000). “A mannose selection system for production of fertile transgenic maize plants from protoplasts.” (1999) Plant Cell Reports 19(7): 654-660.

ß-Gal and ß-GlcU Chemiluminescent Substrates

The lacZ ß-Galactosidase and GUS ß-Glucuronidase reporter genes continue to be among the most popular marker genes in mammalian and plant cell systems, respectively.  Michigan Diagnostics and Marker Gene are working together to develop and market new chemiluminescent substrates based on stabilization using the tricyclo[7.3.1.0^2,7]tridec-2,7-ene] nucleus.  These new, ultra-sensitive lacZ and GUS substrates will be featured in our new catalog for use with transgenic mammalian, bacterial, plant and yeast cell analyses. See the references below for more information about their use.

• Giri, B.P., Giri, K. W., Toben, N. E, and Toben, H. R., “A novel family of chemiluminescent 1,2-dioxetanes”, AACC meeting 2000.
•   Lamkin, M. S., Shinefeld, L., Toben, H. R., Giri, B. P., ”Improved substrate for the detection of secreted placental alkaline phosphatase reporter enzyme” Biolum. Chemilum., April 2002.
  
  

New Catalog in the Mail.

The 2003-2004 edition of the Marker Gene catalog is being mailed out in January.  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.
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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. 

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