title: Identification of differentially expressed genes in anagen dermal papilla by suppression subtractive hybridization.
abstract: BACKGROUND
  We constructed a cDNA subtractive library of dermal papilla cells (DPCs) in anagen with suppression subtractive hybridization (SSH) technique and clone differentially expressed genes related to DPCs in anagen.
  
  
  METHODS
  Total mRNA was isolated from DPCs of anagen and telogen follicles. Moreover, single-strand (ss) and double-strand (ds) cDNAs were synthesized in turn using SMART PCR cDNA synthesis technology. ds cDNAs then were digested with Rsa I and divided into two groups, and ligated to the specific adaptor 1 and adaptor 2R, respectively. After cDNAs were hybridized with each other twice and underwent two rounds of nested PCR. PCR products were ligated with arms of T/A plasmid vectors to set up the subtractive library. Selected clones were demonstrated by reverse Northern blot and sequenced. The acquired sequence data were aligned against the Genbank nucleotide database.
  
  
  RESULTS
  cDNA subtractive library of DPCs in anagen follicles was set up successfully with high subtractive efficiency. Thirty-five genes were identified in this study with 22 known functional genes and 13 unknown functional genes.
  
  
  CONCLUSIONS
  All results confirm the effectiveness and sensitivity of SSH in detecting differentially expressed genes from a small amount of clinical samples. Information about such alterations in gene expression could be useful for elucidating the genetic events in hair follicle growth regulation.
authors.affiliation: Department of Dermatology, Southwest Hospital of Third Military Medical University, Chongqing 400038, China.
authors.forename: Xi-chuan
authors.initials: XC
authors.lastname: Yang
authors.forename: Fei
authors.initials: F
authors.lastname: Hao
authors.forename: Zhi-qiang
authors.initials: ZQ
authors.lastname: Song
authors.forename: Bo
authors.initials: B
authors.lastname: Cheng
authors.forename: Wei-bing
authors.initials: WB
authors.lastname: Yang
authors.forename: Bai-yu
authors.initials: BY
authors.lastname: Zhong
authors.forename: Ming-ming
authors.initials: MM
authors.lastname: Xiang
mesh_terms: D000328:Adult; D000506:Alopecia Areata / Q000235:genetics*; D018076:DNA, Complementary / Q000032:analysis*; D005260:Female; D015723:Gene Library; D006197:Hair; D018859:Hair Follicle / Q000737:chemistry*; D006801:Humans; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction
chemical_list: D018076:DNA, Complementary