人肝癌细胞cDNA文库的构建及鉴定

AIM To construct human hepatocarcinoma cell cDNA library. METHODS Total RNA and purified mRNA were extracted from human hepatocarcinoma cell line HHCC. The single strand and double strand of cDNA were synthesized through reverse transcription. The cDNA fragments, smaller than 500 bp, were removed and the remaining cDNAs were connected with Eco RI adaptors and then combined with the right and left arms of dephophrylated λgt11 Eco RI. The recombinant cDNAs were packaged in vitro , and then a small portion of packaged phage was used to infect E.coli Y1090 for titration. RESULTS The HHCC cell line cDNA library consisting of 1.2×10 6 recombinant bacteriophages was constructed. The average exogenous insert of the recombinants was about 1.5 kb. CONCLUSION The constructed cDNA library can be used to screen target clones.