Cloning and characterization of a novel DNase gene from Trichogramma pretiosum.

Lijuan Yu, Yi Feng, Suxiang Zheng, Jie Li, Ruocen Liu, Dayong Jin

Protein expression and purification 2021 Sep

filter terms:

DNase is a powerful tool for a series of molecular biology applications. Developing a strategy for large-scale production of DNase with high purity and activity is critical for scientific research. In this study, a previously uncharacterized gene with nuclease activity was found in Trichogramma pretiosum genome. Pichia pastoris GS115 was preferred as the host to overcome the issues related to prokaryotic expression. Under the optimal conditions, the activity of T. pretiosum DNase (Tp-DNase) reached 1940 U/mL of culture supernatant in fed-batch fermentation. Using ion-exchange chromatography and adsorption chromatography, Tp-DNase was produced with a purity of >99% and molecular weight of 45 kDa. In vitro DNA degradation experiments showed that Tp-DNase could effectively degrade dsDNA, and its activity was slightly higher than that of bovine pancreas DNase I under the same conditions. Moreover, Tp-DNase can be used to eliminate nucleic acid contamination and improve the accuracy of nucleic acid detection. Copyright © 2021 Elsevier Inc. All rights reserved.

Citation

Lijuan Yu, Yi Feng, Suxiang Zheng, Jie Li, Ruocen Liu, Dayong Jin. Cloning and characterization of a novel DNase gene from Trichogramma pretiosum. Protein expression and purification. 2021 Sep;185:105896