Identification of novel and known oocyte-specific genes using complementary DNA subtraction and microarray analysis in three different species

Biol Reprod. 2005 Jul;73(1):63-71. doi: 10.1095/biolreprod.104.037069. Epub 2005 Mar 2.


The main objective of the present study was to identify novel oocyte-specific genes in three different species: bovine, mouse, and Xenopus laevis. To achieve this goal, two powerful technologies were combined: a polymerase chain reaction (PCR)-based cDNA subtraction, and cDNA microarrays. Three subtractive libraries consisting of 3456 clones were established and enriched for oocyte-specific transcripts. Sequencing analysis of the positive insert-containing clones resulted in the following classification: 53% of the clones corresponded to known cDNAs, 26% were classified as uncharacterized cDNAs, and a final 9% were classified as novel sequences. All these clones were used for cDNA microarray preparation. Results from these microarray analyses revealed that in addition to already known oocyte-specific genes, such as GDF9, BMP15, and ZP, known genes with unknown function in the oocyte were identified, such as a MLF1-interacting protein (MLF1IP), B-cell translocation gene 4 (BTG4), and phosphotyrosine-binding protein (xPTB). Furthermore, 15 novel oocyte-specific genes were validated by reverse transcription-PCR to confirm their preferential expression in the oocyte compared to somatic tissues. The results obtained in the present study confirmed that microarray analysis is a robust technique to identify true positives from the suppressive subtractive hybridization experiment. Furthermore, obtaining oocyte-specific genes from three species simultaneously allowed us to look at important genes that are conserved across species. Further characterization of these novel oocyte-specific genes will lead to a better understanding of the molecular mechanisms related to the unique functions found in the oocyte.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • DNA, Complementary / genetics*
  • Female
  • Gene Expression Regulation
  • Mice
  • Mice, Inbred BALB C
  • Nucleic Acid Hybridization
  • Oligonucleotide Array Sequence Analysis
  • Oocytes / physiology*
  • Ovary / physiology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Species Specificity
  • Xenopus laevis


  • DNA, Complementary