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Reconstruction of a hybrid nucleoside antibiotic gene cluster based on scarless modification of large DNA fragments

Overview of attention for article published in Science China Life Sciences, August 2017
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Title
Reconstruction of a hybrid nucleoside antibiotic gene cluster based on scarless modification of large DNA fragments
Published in
Science China Life Sciences, August 2017
DOI 10.1007/s11427-017-9119-1
Pubmed ID
Authors

Jiming Zhuo, Binbin Ma, Jingjing Xu, Weihong Hu, Jihui Zhang, Huarong Tan, Yuqing Tian

Abstract

Genetic modification of large DNA fragments (gene clusters) is of great importance in synthetic biology and combinatorial biosynthesis as it facilitates rational design and modification of natural products to increase their value and productivity. In this study, we developed a method for scarless and precise modification of large gene clusters by using RecET/RED-mediated polymerase chain reaction (PCR) targeting combined with Gibson assembly. In this strategy, the biosynthetic genes for peptidyl moieties (HPHT) in the nikkomycin biosynthetic gene cluster were replaced with those for carbamoylpolyoxamic acid (CPOAA) from the polyoxin biosynthetic gene cluster to generate a ~40 kb hybrid gene cluster in Escherichia coli with a reusable targeting cassette. The reconstructed cluster was introduced into Streptomyces lividans TK23 for heterologous expression and the expected hybrid antibiotic, polynik A, was obtained and verified. This study provides an efficient strategy for gene cluster reconstruction and modification that could be applied in synthetic biology and combinatory biosynthesis to synthesize novel bioactive metabolites or to improve antibiotic production.

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Mendeley readers

Mendeley readers

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Geographical breakdown

Country Count As %
Unknown 5 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 2 40%
Professor > Associate Professor 1 20%
Student > Doctoral Student 1 20%
Student > Master 1 20%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 2 40%
Computer Science 1 20%
Agricultural and Biological Sciences 1 20%
Unknown 1 20%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 26 August 2017.
All research outputs
#20,444,703
of 22,999,744 outputs
Outputs from Science China Life Sciences
#774
of 1,009 outputs
Outputs of similar age
#277,335
of 317,627 outputs
Outputs of similar age from Science China Life Sciences
#26
of 35 outputs
Altmetric has tracked 22,999,744 research outputs across all sources so far. This one is in the 1st percentile – i.e., 1% of other outputs scored the same or lower than it.
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