title: [Cloning associated genes using microdissection-cDNA PCR-SSH in gastric dysplasia].
abstract: OBJECTIVE
  To construct cDNA subtracted libraries from gastric dysplasia and further screen differentially expressed genes.
  
  
  METHODS
  Relatively pure dysplasia and normal tissue were procured by manual microdissection, and amplified by cDNA-PCR, which was used to carry on for suppression subtractive hybridization (SSH). Subtracted cDNA fragments were linked with vector, cloned, screened, sequenced, and made homologous search. Differentially expressed fragments were verified by dot hybridization.
  
  
  RESULTS
  Two subtracted cDNA libraries were constructed. Among 26 sequenced clones, 15 fragments corresponded to known genes, 3 fragments were known EST and 8 fragments were unknown EST (GenBank BQ164614-BQ164616, BQ291516-BQ291520). Fifteen fragments were verified to be differentially expressed in gastric dysplasia.
  
  
  CONCLUSIONS
  Subtracted cDNA libraries from gastric dysplasia are constructed using combination of microdissection-cDNA PCR and SSH setup in our laboratory. Some fragments have been screened and verified to help to search for novel associated genes with gastric carcinogenesis.
authors.affiliation: Department of Medical Genetics, China Medical University, Shenyang 110001, China.
authors.forename: Dong-mei
authors.initials: DM
authors.lastname: Hao
authors.forename: Xiu-ju
authors.initials: XJ
authors.lastname: Sun
authors.forename: Zhi-hong
authors.initials: ZH
authors.lastname: Zheng
authors.forename: Guang
authors.initials: G
authors.lastname: He
authors.forename: Ming-chao
authors.initials: MC
authors.lastname: Ma
authors.forename: Hui-mian
authors.initials: HM
authors.lastname: Xu
authors.forename: Mei-xian
authors.initials: MX
authors.lastname: Wang
authors.forename: Kai-lai
authors.initials: KL
authors.lastname: Sun
mesh_terms: D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D006801:Humans; D042282:Microdissection; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction; D011230:Precancerous Conditions / Q000235:genetics* / Q000473:pathology; D017422:Sequence Analysis, DNA; D013274:Stomach Neoplasms / Q000235:genetics* / Q000473:pathology
chemical_list: D018076:DNA, Complementary