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  • Title: [Construction and Identification of the cDNA Expression Library for Human Esophageal Cancer Cells].
    Author: Zhang Z, Wu XY, Feng L, Huang SK, Luo MN, Shao S, Zhao XH.
    Journal: Sichuan Da Xue Xue Bao Yi Xue Ban; 2016 Sep; 47(5):786-789. PubMed ID: 28598100.
    Abstract:
    OBJECTIVES: To construct a cDNA phage expression library for human esophageal cancer cells. METHODS: After the total RNA were obtained from esophageal cancer cells, the mRNA were separated with magnetic beads adsorption method, and the single-strand and double-strand cDNA were synthesized through reverse transcription. With the undesirable cDNA fragments removed, the remaining cDNA (linked withEcoR1 aptamer and phosphorylated its 5'end) combined with the carrier of T7 Select10-3b. The recombinant phage were packaged in vitro for preliminary cDNA library. PCR was used to identify the size of inserted cDNA. RESULTS: The constructed original cDNA phage expression library for human esophageal cancer cells was consisted of 2.01×10⁶ pfu/mL bacteriophages with a recombination rate of 100%. The length of the inserted cDNA fragments were range from 300 bp to 1 500 bp. CONCLUSIONS: The cDNA phage expression library of human esophageal cell is successfully constructed to meet the currently recognized standards, and can be well used to screen cDNA-cloned genes of human esophageal cancer antigens by serological analysis of recombinantly expressed cDNA clone (SEREX).
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