title: [Construction and identification of the expression library of album pollen allergens cDNA].
abstract: OBJECTIVE
  To construct and identify the express library of album pollen allergens cDNA.
  
  
  METHODS
  Total RNA were extracted from the album pollen with TRIzol reagent and the mRNA was isolate for the amplify followed. A double stranded cDNA (ds cDNA) was synthesized using primers containing Xho I and Poly(dT) sequence by ZAP Express®cDNA synthesis kit. The ds cDNA was modified and purified by gel chromatography, and then the cDNA fragment with the length of more than 400 bp containing sticky ends was obtained. The cDNA fragment was ligated with Uni-ZAP XR vector and subsequently treated with in vitro packaging using phage by ZAP-cDNA express GigapackIII Gold cloning kit. The express library of album pollen cDNA was constructed by in vitro packaging. The recombination rate and the lengths of fragments inserted of the cDNA library were detected by polymerase chain reaction.
  
  
  RESULTS
  The titer and the recombination rate of cDNA expression library constructed were 9.7×10(5) and 100%, respectively. The capacity of the library was 4.85 Pfu. The average length of cDNA fragments inserted was about 1.0 kb.
  
  
  CONCLUSIONS
  Based on the capacity of cDNA expression library constructed and the length of cDNA insertion fragments, the cDNA expression library constructed is qualified to screening target cDNA clone, laying the foundation for preparation of gene recombinant allergen pollen vaccine.
authors.affiliation: Department of Respiration, Second Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an 710004, China. zhangjie@mail.xjtu.edu.cn
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Xiu-zhen
authors.initials: XZ
authors.lastname: Sun
authors.forename: Hong
authors.initials: H
authors.lastname: Yan
authors.forename: Ni
authors.initials: N
authors.lastname: Zhang
authors.forename: Xiang-li
authors.initials: XL
authors.lastname: Feng
mesh_terms: D000485:Allergens / Q000737:chemistry / Q000235:genetics*; D027463:Chenopodium album / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D015723:Gene Library; D011058:Pollen / Q000235:genetics* / Q000276:immunology
chemical_list: D000485:Allergens