title: Identification of differentially expressed genes in murine embryos at the blastocyst stage using annealing control primer system.
abstract: The identification of embryo-specific genes would provide insights into early embryonic development. However, the current methods employed to identify the genes that are expressed at a specific developmental stage are labor intensive and suffer from high rates of false positives. Here we employed a new and accurate reverse transcription-polymerase chain reaction (RT-PCR) method that involves annealing control primers (ACPs) to identify the genes that are specifically or prominently expressed in mouse blastocysts compared to 4-cell stage embryos. Using 120 ACPs, we identified and sequenced 74 of these differentially expressed genes (DEGs). Basic Local Alignment Search Tool (BLAST) searches revealed that 53 were known genes, 9 encoded ribosomal proteins, and 12 were unknown genes. Of the known genes, 14 were selected and further characterized using real-time quantitative PCR to assess their stage-specific expression in mouse embryos. This analysis suggests that the ACP system is a very good method for the identification of stage-specific genes in small numbers of mouse embryos. Further analysis of the differentially expressed blastocyst genes we have identified will provide insights into the molecular basis of preimplantation development.
authors.affiliation: Department of Animal Science, Chungbuk National University, Gaesin-dong, Cheongju, Chungbuk, 361-763, South Korea.
authors.forename: Xiang-Shun
authors.initials: XS
authors.lastname: Cui
authors.forename: Mi-Ra
authors.initials: MR
authors.lastname: Shin
authors.forename: Kyung-Ah
authors.initials: KA
authors.lastname: Lee
authors.forename: Nam-Hyung
authors.initials: NH
authors.lastname: Kim
mesh_terms: D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000378:metabolism*; D019295:Computational Biology; D017931:DNA Primers; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D005796:Genes / Q000235:genetics*; D016014:Linear Models; D051379:Mice; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
chemical_list: D017931:DNA Primers