title: [Construction and screening of cDNA library of osteosarcoma cells].
abstract: OBJECTIVE
  To construct a cDNA library of osteosarcoma SAOS-2 cells and screen the proteins interacting with the bone morphogenetic protein-2 (BMP-2) to explore the role of BMP-2 in the development of prostate cancer.
  
  
  METHODS
  The total RNA was extracted from SAOS-2 cells, from which poly(A)(+)RNA was purified to generate the cDNA using RT-PCR with primer SMARTIII and primer CDSIIIoligo(dT). The library was constructed and screened by cotransformation of the double-stranded cDNA (dscDNA) and linearized pGADT7-Rec with the bait plasmid to the yeast AH109. The proteins interacting with BMP-2 were screened by AH109 mating with the bait strain Y187, and the interaction was confirmed using far-Western blot analysis.
  
  
  RESULTS
  The cDNA library of human osteosarcoma SAOS-2 cells was constructed, which was characterized by high diversity and sufficient capacity. The dscDNA size range was between 250-5000 bp, with a cotransformation efficiency of 4.3 x 10(5) and recombination efficiency of 1.9 x 10(6), and 4.3 x 10(5) clones were screened. The mating efficiency was 32% and 1.0 x 10(6) clones were screened by the bait of BMP-2, and 4 positive clones was obtained and sequenced.
  
  
  CONCLUSIONS
  The diversity and capacity of the cDNA library meet the needs for screening genes related to prostate cancer.
authors.affiliation: General Hospital of Chengdu Command, Chengdu 610083, China. deng shaolin@medmail.com.cn
authors.forename: Shao-lin
authors.initials: SL
authors.lastname: Deng
authors.forename: Yi
authors.initials: Y
authors.lastname: Quan
authors.forename: Xian-ming
authors.initials: XM
authors.lastname: Pan
authors.forename: Bo
authors.initials: B
authors.lastname: Qü
mesh_terms: D055396:Bone Morphogenetic Protein 2 / Q000235:genetics; D045744:Cell Line, Tumor; D015723:Gene Library; D006801:Humans; D008297:Male; D012516:Osteosarcoma / Q000235:genetics*; D011471:Prostatic Neoplasms / Q000235:genetics; D012334:RNA, Neoplasm / Q000235:genetics; D020798:Two-Hybrid System Techniques
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D012334:RNA, Neoplasm