Construction and characterization of K562 cell cDNA bank

Gang Chen, Wanggang Zhang, Jie Fu, Fuyang Li, Xinping Liu, Libo Yao

Research output: Contribution to journalArticle

Abstract

Objective: To construct a human K562 cell cDNA expression bank. Methods: Total RNA and purified mRNA were extracted from leukemia cell line K562. First and second strand of cDNA were synthesized through reverse transcription. After blunting the cDNA termini, the cDNA fragments were connected with EcoR I adapters, and the end of EcoR I adapters was phosphorylated. The cDNA smaller than 400 bp were removed by Sephacryl-S400 spin column, and the remaining were ligated with the dephosphorylated arms of λgt11. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli Y1090 for titration. Results: The cDNA bank consisting of 1.02 × 106 recombinant bacteriophages was constructed. The average exogenous insertion of the recombinants was about 1.3 kb. Conclusion: The constructed cDNA bank can be used to screen target clones.

Original languageEnglish (US)
Pages (from-to)109-111
Number of pages3
JournalJournal of Xi'an Jiaotong University (Medical Sciences)
Volume24
Issue number2
StatePublished - Apr 1 2003

Keywords

  • K562 cell line
  • Leukemia
  • cDNA bank

ASJC Scopus subject areas

  • Clinical Biochemistry

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