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[DNA family shuffling of tissue-type plasminogen activator genes]

Qing-sheng Huang1, Jing Xu, Jun-jie Xu

  • 1Institute of Microbiology and Epidemiology, AMMS, Beijing 100071, China.

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi = Chinese Journal of Cellular and Molecular Immunology
|May 11, 2004
PubMed
Abstract

Insights

DNA shuffling successfully identified a tissue plasminogen activator (tPA) variant with fourfold higher activity. Another variant showed normal activity despite an 88-amino-acid deletion, paving the way for enhanced tPA screening.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Protein Engineering

Context:

  • Tissue plasminogen activator (tPA) is crucial for fibrinolysis.
  • Enhancing tPA activity is a key goal in thrombolytic therapy.
  • DNA shuffling offers a powerful method for protein engineering.

Purpose:

  • To explore DNA shuffling for screening higher-activity tissue plasminogen activator (tPA).
  • To generate and evaluate a diverse library of tPA variants.

Summary:

  • Human, rat, and rhesus monkey tPA cDNAs were used for DNA family shuffling.
  • A library of shuffled tPA variants was expressed in CHO cells and screened.
  • Two clones, t9 and t17, were selected: t9 exhibited 4x higher activity, while t17 showed normal activity with an 88-amino-acid deletion.

Impact:

  • This study demonstrates the potential of DNA shuffling to discover improved tPA variants.
  • The findings lay the groundwork for future research in screening for enhanced tPA activity.
  • Identified variants offer insights into structure-activity relationships of tPA.

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