Screening of phage displayed cDNA library of human liver against methotrexate on agrose gel matrix

China Biotechnology

2007, Vol. 27

Issue (10): 70-74 DOI:

Screening of phage displayed cDNA library of human liver against methotrexate on agrose gel matrix

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Abstract

The screening of the specific interaction between proteins displayed on phage displayed human liver cDNA library with methotrexate was carried out. Methods: With methotrexate immobilized on agarose gel, the affinity selection process of "binding-eluting-amplifying" was performed for five rounds to enrich the specific phage clones. PCR was performed to monitor the selection process, and the amplification products were sequenced and searched in GenBank by BLAST to find the obtained target proteins with high similarity. Results: The PCR products of the selected phage clones were proven with identical sequence with PI-3K related kinase SMG-1 isoform 1. Conclusions: The screening of specific proteins by using affinity selection of phage displayed cDNA library with small-molecule drug immobilized on agarose was an efficient and convenient method. It might be good to search for the candidate targets of the drug molecules, and might provide clues for the further studies of drug mechanism and toxocity.

Key words： phage display human liver cDNA methotrexate interaction

Received: 25 June 2007 Published: 25 October 2007

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Cite this article:

. Screening of phage displayed cDNA library of human liver against methotrexate on agrose gel matrix. China Biotechnology, 2007, 27(10): 70-74.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2007/V27/I10/70

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