Vol.2 No.2 (April 2012)
Construction of a T7 Phage Display cDNA Library from Lung Cancer of Smokers
钟理 ：河北大学生命科学学院，保定 College of Osteopathic Medicine, Pomona, USA
Objective: To screen tumor markers of smokers, the T7 phage display cDNA library from the lung cancer of smokers was constructed. Methods: The mRNA was isolated from total RNA extracted from the lung cancer of smokers by RNeasy Mini Kit, and was used to synthesize double strain (ds) cDNA by the reverse transcription. Then the directional EcoRI/HindIII linkers were liquated into the ends of ds cDNA and the ds cDNA was further digested with EcoRI and HindIII, which resulted in ds cDNA with EcoRI and HindIII ends. The digested ds cDNA fragments longer than 300 bp in length were fractionated by Mini Column, and then liquated into the T7 Select 10-3b vertor with EcoRI and HindIII ends. After packaging in vitro, the T7 Select 10-3b vertor was tansformed into BLT5615 to construct the T7 phage display cDNA library. Results: Analysis showed that the library contained 1.35 × 106 clones and the titer of the applied library was 4.4 × 1010 pfu/ml. The PCR identification results of 100 clones picked at random showed that 97% clones were recombinant and 91% of recombinant clones contained cDNA fragments longer than 300 bp in length. Conclusion: A T7 phage display cDNA library from the lung cancer of smokers is successfully constructed.
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