Differential screening of a subtracted cDNA library: a method to search for genes preferentially expressed in multiple tissues

BioTechniques
H JinC C Huang

Abstract

We have developed a new strategy for differential screening of genes that are expressed in two or more tissues, and have used it to identify genes that are preferentially expressed in both testis and ovary. In this approach, testis-specific cDNAs were first generated using the suppression subtractive hybridization technique, cloned and arrayed in microplates. The inserts from the subtracted testis-specific library were amplified by PCR and spotted on filters. The dot blots were screened by hybridization using either testis- or ovary-specific subtracted cDNA mixtures as probes. By screening about 2000 putative clones from the subtracted testis-specific library, we identified 14 candidate genes that hybridized to both cDNA probes. Northern blot analysis showed that 12 clones were preferentially or exclusively expressed in testis and ovary. Nucleotide sequence analysis indicated that three of the identified clones represented cDNAs from novel genes.

Citations

Feb 9, 1999·Prenatal Diagnosis·E Pergament, M Fiddler
May 11, 2004·Journal of Experimental Botany·Masaru KobayashiToru Matoh
Jul 19, 2005·Proceedings. Biological Sciences·Jeffrey J M PereboomAndrew F G Bourke
Jul 15, 2005·Annals of Periodontology·Denis RebrikovLuda Diatchenko
Sep 1, 2004·BioTechniques·Jeffrey Scott Coker, Eric Davies
Aug 13, 2004·Brain Research. Molecular Brain Research·E YakubovE Yavin
Jun 2, 2004·Molecular Carcinogenesis·Jin He, Lung-Ji Chang

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