%0 Journal Article %T Construction of a T7 phage display cDNA library from lung of Microtus |fortis %D 2013 %R %J Chinese Journal of Schistosomiasis Control %P 252-256 %V 19 %N 4 %X

  Objective  To construct a T7 phage display cDNA library from the lung of Microtus
fortis for further screening the schistosomiasis-resistence-related genes of Microtus fortis. Methods
  mRNA was isolated from total RNA extracted from the lungs of Microtus fortis by TRIzol
reagent, and was used to synthesize double strain cDNA by the reverse transcription. Then the
double strain cDNA was given with EcoR I  and Hind I[ adhering ends by ligation with the direc-
tional EcoR I /Hind II linkers and digestion with EcoR i  and Hind  m.  The double strain cDNA
fragments longer than 300 bp in length were fractionated by the Mini Column, and ligated into the
T7 Select l0-3b vector with EcoR I and Hind m adhering ends. After packaging in vitro, the re-
combinant T7 Select l0-3b was transformed into BLT5403 to construct a T7 phage display cDNA li-
brary.  Results   The library constructed here contained l. 5 Xl06 clones and the titer of the amplied
library was l. 1X101z pfu/ml.  The PCR identification results of 100 clones picked at random showed
that 91% clones were recombinant and 90% of recombinant clones contained cDNA fragments
longer than 300 bp in length. Conclusion  A T7 phage display cDNA library from the lung of M/-
crotus fortis is successfully constructed.
   

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