Characterization and application of a thermophilic Argonaute from archaeon Thermococcus thioreducens

被引：19
作者：
Fang, Mengjun [1 ,2 ]
Xu, Zhipeng [3 ]
Huang, Di [1 ,2 ]
Naeem, Muhammad [1 ,2 ]
Zhu, Xiangcheng [4 ]
Xu, Zhinan [1 ,2 ]
机构：
[1] Zhejiang Univ, Coll Chem & Biol Engn, Key Lab Biomass Chem Engn, Minist Educ, Hangzhou, Peoples R China
[2] Zhejiang Univ, Inst Biol Engn, Coll Chem & Biol Engn, Hangzhou 310027, Peoples R China
[3] Zhejiang Univ, Affiliated Hosp 1, Sch Med, Hangzhou, Peoples R China
[4] Cent South Univ, Xiangya Int Acad Translat Med, Changsha, Hunan, Peoples R China
来源：
BIOTECHNOLOGY AND BIOENGINEERING | 2022年 / 119卷 / 09期
基金：
中国国家自然科学基金;
关键词：
DNA detection; programmable DNA endonuclease; prokaryotic Argonaute protein; DNA; CLEAVAGE;
D O I：
10.1002/bit.28153
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
Prokaryotic Argonaute proteins (pAgos) play an important role in host defense against invading genetic elements. The functional diversities make pAgos very promising in development of novel nucleic acid manipulation tools and attract increasing attentions. Here, we reported the in vitro characterization of an Argonaute protein from archaeon Thermococcus thioreducens (TtrAgo) and its example of application in hepatitis B virus DNA detection. The results showed that TtrAgo functions as a programmable DNA endonuclease by utilizing both short 5 '-phosphorylated and 5 '-hydroxylated single-stranded DNA guides, and presents high efficiency and accuracy at optimal temperatures ranging from 75 degrees C to 95 degrees C. In addition, TtrAgo also possesses stepwise cleavage activity like PfAgo (Pyrococcus furiosus) and chopping activity toward double-stranded DNA similar to MjAgo (Methanocaldococcus jannaschii). This study increases our understanding of pAgos and expands the Ago-based DNA detection toolbox.
引用
收藏
页码：2388 / 2398
页数：11
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