Engineering of the Fluorescent‐Energy‐Conversion Arm of Phi29 DNA Packaging Motor for Single‐Molecule Studies |
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Authors: | Tae Jin Lee Hui Zhang Chun‐Li Chang Cagri Savran Peixuan Guo |
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Affiliation: | 1. Department of Biomedical Engineering The Vontz Center for Molecular Studies 3125 Eden Avenue, Room 1301 College of Engineering and College of Medicine University of Cincinnati Cincinnati, OH 45267 (USA);2. School of Electrical and Computer Engineering School of Mechanical Engineering Weldon School of Biomedical Engineering Purdue University West Lafayette, IN 47907 (USA) |
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Abstract: | The bacteriophage phi29 DNA packaging motor contains a protein core with a central channel comprising twelve copies of re‐engineered gp10 protein geared by six copies of packaging RNA (pRNA) and a DNA packaging protein gp16 with unknown copies. Incorporation of this nanomotor into a nanodevice would be beneficial for many applications. To this end, extension and modification of the motor components are necessary for the linkage of this motor to other nanomachines. Here the re‐engineering of the motor DNA packaging protein gp16 by extending its length and doubling its size using a fusion protein technique is reported. The modified motor integrated with the eGFP‐gp16 maintains the ability to convert the chemical energy from adenosine triphosphate (ATP) hydrolysis to mechanical motion and package DNA. The resulting DNA‐filled capsid is subsequently converted into an infectious virion. The extended part of the gp16 arm is a fluorescent protein eGFP, which serves as a marker for tracking the motor in single‐molecule studies. The activity of the re‐engineered motor with eGFP‐gp16 is also observed directly with a bright‐field microscope via its ability to transport a 2‐µm‐sized cargo bound to the DNA. |
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Keywords: | DNA nanobiotechnology nanoparticles packaging motors |
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