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A Histone-Like Protein Induces Plasmid DNA to Form Liquid Crystals in Vitro and Gene Compaction in Vivo
Key Laboratory of Solid Waste Treatment and Resource Recycle & Fundamental Science on Nuclear Waste and Environmental Security Laboratory, Southwest University of Science and Technology, Mianyang 621010, Sichuan, China
School of Environment and Resource, Southwest University of Science and Technology, Mianyang 621010, Sichuan, China
* Authors to whom correspondence should be addressed.
Received: 30 October 2013; in revised form: 17 November 2013 / Accepted: 21 November 2013 / Published: 6 December 2013
Abstract: The liquid crystalline state is a universal phenomenon involving the formation of an ordered structure via a self-assembly process that has attracted attention from numerous scientists. In this study, the dinoflagellate histone-like protein HCcp3 is shown to induce super-coiled pUC18 plasmid DNA to enter a liquid crystalline state in vitro, and the role of HCcp3 in gene condensation in vivo is also presented. The plasmid DNA (pDNA)-HCcp3 complex formed birefringent spherical particles with a semi-crystalline selected area electronic diffraction (SAED) pattern. Circular dichroism (CD) titrations of pDNA and HCcp3 were performed. Without HCcp3, pUC18 showed the characteristic B conformation. As the HCcp3 concentration increased, the 273 nm band sharply shifted to 282 nm. When the HCcp3 concentration became high, the base pair (bp)/dimer ratio fell below 42/1, and the CD spectra of the pDNA-HCcp3 complexes became similar to that of dehydrated A-form DNA. Microscopy results showed that HCcp3 compacted the super-coiled gene into a condensed state and that inclusion bodies were formed. Our results indicated that HCcp3 has significant roles in gene condensation both in vitro and in histone-less eukaryotes in vivo. The present study indicates that HCcp3 has great potential for applications in non-viral gene delivery systems, where HCcp3 may compact genetic material to form liquid crystals.
Keywords: liquid crystal; gene condensation; plasmid; histone-like protein; non-viral gene delivery
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MDPI and ACS Style
Sun, S.; Liu, M.; Dong, F.; Fan, S.; Yao, Y. A Histone-Like Protein Induces Plasmid DNA to Form Liquid Crystals in Vitro and Gene Compaction in Vivo. Int. J. Mol. Sci. 2013, 14, 23842-23857.
Sun S, Liu M, Dong F, Fan S, Yao Y. A Histone-Like Protein Induces Plasmid DNA to Form Liquid Crystals in Vitro and Gene Compaction in Vivo. International Journal of Molecular Sciences. 2013; 14(12):23842-23857.
Sun, Shiyong; Liu, Mingxue; Dong, Faqin; Fan, Shenglan; Yao, Yanchen. 2013. "A Histone-Like Protein Induces Plasmid DNA to Form Liquid Crystals in Vitro and Gene Compaction in Vivo." Int. J. Mol. Sci. 14, no. 12: 23842-23857.