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

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Jin, H; Cheng, X; Diatchenko, L; Siebert, PD; Huang, CC

Published Date

  • December 1997

Published In

Volume / Issue

  • 23 / 6

Start / End Page

  • 1084 - 1086

PubMed ID

  • 9421640

International Standard Serial Number (ISSN)

  • 0736-6205

Digital Object Identifier (DOI)

  • 10.2144/97236st05


  • eng

Conference Location

  • England