Development of a 950-gene DNA array for examining gene expression patterns in mouse testis

Rockett, Christopher, Brian, Krawetz, Hughes, Hee, Oudes, Dix

Description

Over the past five years, interest in and use of DNA array technology has increased dramatically, and there has been a surge in demand for different types of arrays. Although manufacturers offer a number of pre-made arrays, these are generally of utilitarian design and often cannot accommodate the specific requirements of focused research, such as a particular set of genes from a particular tissue. We found that suppliers did not provide an array to suit our particular interest in testicular toxicology, and therefore elected to design and produce our own. Results We describe the procedures used by members of the US Environmental Protection Agency MicroArray Consortium (EPAMAC) to produce a mouse testis expression array on both filter and glass-slide formats. The approaches used in the selection and assembly of a pertinent, nonredundant list of testis-expressed genes are detailed. Hybridization of the filter arrays with normal and bromochloroacetic acid-treated mouse testicular RNAs demonstrated that all the selected genes on the array were expressed in mouse testes. Conclusion We have assembled two lists of mouse (950) and human (960) genes expressed in the mouse and/or human adult testis, essentially all of which are available as sequence-verified clones from public sources. Of these, 764 are homologous and will therefore enable close comparison of gene expression between murine models and human clinical testicular samples.

Informations

• Publié par: biomed
• Publié le: 01 janvier 2001
• Nombre de lectures: 1
• Langue: English
• Poids de l'ouvrage: 1 Mo

Extrait

http://genomebiology.com/2001/2/4/research/0014.1

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Published: 22 March 2001 GenomeBiology2001,2(4):research0014.1–0014.9 The electronic version of this article is the complete one and can be found online at http://genomebiology.com/2001/2/4/research/0014 © 2001 Rockettet al., licensee BioMed Central Ltd (Print ISSN 14656906; Online ISSN 14656914)

Received: 17 November 2000 Revised: 27 December 2000 Accepted: 31 January 2001

Abstract Background:Over the past five years, interest in and use of DNA array technology has increased dramatically, and there has been a surge in demand for different types of arrays. Although manufacturers offer a number of premade arrays, these are generally of utilitarian design and often cannot accommodate the specific requirements of focused research, such as a particular set of genes from a particular tissue. We found that suppliers did not provide an array to suit our particular interest in testicular toxicology, and therefore elected to design and produce our own. Results:We describe the procedures used by members of the US Environmental Protection Agency MicroArray Consortium (EPAMAC) to produce a mouse testis expression array on both filter and glassslide formats. The approaches used in the selection and assembly of a pertinent, nonredundant list of testisexpressed genes are detailed. Hybridization of the filter arrays with normal and bromochloroacetic acidtreated mouse testicular RNAs demonstrated that all the selected genes on the array were expressed in mouse testes.

Conclusion:We have assembled two lists of mouse (950) and human (960) genes expressed in the mouse and/or human adult testis, essentially all of which are available as sequenceverified clones from public sources. Of these, 764 are homologous and will therefore enable close comparison of gene expression between murine models and human clinical testicular samples.

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