Construction and characterization of osteosarcoma 9901 cell cDNA library

AIM: To construct human osteosarcoma 9901 cell cDNA expression library for screening osteosarcoma-specific antigens. METHODS: Total RNA was extracted from human osteosarcoma cell line 9901 and mRNA was purified. cDNA was synthesized by reverse transcription, and cDNA fragments larger than 400 bp were ligated with dephosphorylated arms of Agt11. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E coli Y1090 for titration. The size of cDNA inserts and the diversity of library were detected by PCR. RESULTS: The osteosarcoma 9901 cell line cDNA library consisting of 1.5 × 106 recombinant bacteriophages was constructed. The average length of exogenous inserts in the recombinants was about 1.4 kb. CONCLUSION: The cDNA library reported herein is suitable for screening osteosarcoma-specific antigens.