Identification of Differentially Expressed Genes in Anagen Dermal Papilla by Suppression Subtractive Hybridization
- VernacularTitle:抑制性消减杂交筛选生长期毛乳头细胞差异表达基因
- Author:
Xichuan YANG
;
Fei HAO
;
Weibing YANG
;
Zhiqiang SONG
;
Bo CHENG
;
Baiyu ZHONG
;
Mingming XIANG
- Publication Type:Journal Article
- Keywords:
Hair follicle;
Alopecia areata;
Gene library;
Suppression subtractive hybridization
- From:
Chinese Journal of Dermatology
2003;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
Objectives To construct a cDNA subtractive library of dermal papilla cells (DPCs) in anagen with suppression subtractive hybridization (SSH) and clone differentially expressed genes related to DPCs in anagen. Methods Total RNA was isolated from DPC of anagen and telogen follicles. Then ds cDNAs were synthesized in turn using SMART cDNA synthesis technique. After cDNAs from anagen and telogen follicle DPCs 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 verified by reverse Nothern blot and DNA sequencing, and the acquired sequences were analyzed for homology based on Genbank nucleotide database. Results cDNA subtractive library of DPCs in anagen follicle was set up successfully with high subtractive efficiency. Thirty-five genes were identified with 22 known functional genes and 13 unknown functional genes. Conclusions These results demonstrate 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 might be useful for elucidating the genetic events in hair follicle growth regulation.
