title: [Construction and expression identification of human secreted apoptosis-related protein 1 gene yeast two-hybrid bait vector].
abstract: OBJECTIVE
  To construct human secreted apoptosis-related protein 1 (SARP1) gene yeast two-hybrid bait vector so as to study the biological functions of the SARP1 gene in the scar tissue.
  
  
  METHODS
  The target gene from SARP1 gene full-length DNA segment was amplified by PCR, the upstream and downstream primers of the SARP1 gene with restriction enzymes Nde I and Sal I were designed. pGBKT7-SARP1 recombination plasmid was constructed by ligating the vector and the PCR production and identified by PCR and sequencing. Further more, pGBKT7-SARP1 was transformed into competent AH109 which contained kanamycin for selecting positive clones and screened the positive clony on the plate of SD/-Trp. The toxicity and transcriptional activation were tested by the phenotype assay.
  
  
  RESULTS
  SARP1 was amplified and cloned into pGBKT7 successfully, SARP1 gene sequence in recombinant plasmid pGBKT7-SARP1 was verified by gel electrophoresis and DNA sequencing analysis. The sequence of inserted SARP1 gene was the same as the corresponding sequence found in GenBank database. The recombinant pGBKT7-SARP1 plasmids and empty pGBKT7 vector could form white colonies on SD/-Trp plates and none could survive on SD/-Leu plates.
  
  
  CONCLUSIONS
  The recombinant pGBKT7-SARP1 gene yeast two-hybrid bait vector is successfully constructed.
authors: Department of Medical Cosmetology, the Affiliated Hospital ofXi'an Medical University, Xi'an Shaanxi, 710077, P.R.China.
authors: Wei
authors: W
authors: Zhang
authors: Guangzhao
authors: G
authors: He
authors: Bing
authors: B
authors: Ma
mesh_terms: D017209:Apoptosis; D002921:Cicatrix / Q000235:genetics / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics; D017422:Sequence Analysis, DNA; D020798:Two-Hybrid System Techniques; D000094742:Secreted Frizzled-Related Proteins
chemical_list: D008565:Membrane Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; C109618:SFRP2 protein, human; D000094742:Secreted Frizzled-Related Proteins