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Single DNA Condensation Induced by Hexammine Cobalt with Molecular Combing
Gao-ming Hu,Yu Lin,Shi-yong Ran,Yan-wei Wang,Guang-can Yang*
Author NameAffiliationE-mail
Gao-ming Hu College of Physics and Electronic Information Engineering, Wenzhou University, Wenzhou 325035, China  
Yu Lin College of Physics and Electronic Information Engineering, Wenzhou University, Wenzhou 325035, China  
Shi-yong Ran College of Physics and Electronic Information Engineering, Wenzhou University, Wenzhou 325035, China  
Yan-wei Wang College of Physics and Electronic Information Engineering, Wenzhou University, Wenzhou 325035, China  
Guang-can Yang* College of Physics and Electronic Information Engineering, Wenzhou University, Wenzhou 325035, China yanggc@wzu.edu.cn 
Abstract:
We investigated the interaction between DNA and hexammine cobalt III [Co(NH3)6]3+ by a simple molecular combing method and dynamic light scattering. The average extension of ?λ-DNA-YOYO-1 complex is found to be 20.9 μm, about 30% longer than the contour length of the DNA in TE buffer (10 mmol/L Tris, 1 mmol/L EDTA, pH=8.0), due to bis-intercalation of YOYO-1. A multivalent cation, hexammine cobalt, is used for DNA condensation. We find that the length of DNA-[Co(NH3)6]3+ complexes decrease from 20.9 μm to 5.9 μm as the concentration of the [Co(NH3)6]3+ vary from 0 to 3 μmol/L. This observation provides a direct visualization of single DNA condensation induced by hexammine cobalt. The results from the molecular combing studies are supported by dynamic light scattering investigation, where the average hydrodynamic radius of the DNA complex decreases from 203.8 nm to 39.26 nm under the same conditions. It shows that the molecular combing method is feasible for quantitative conformation characterization of single bio-macromolecules.
Key words:  Molecular combing, Hexammine cobalt, Condensation, Dynamic light scatter-ing, Fluorescent microscopy
FundProject:
三氯六氨络合钴导致DNA凝聚的分子梳研究
胡高铭,林瑜,冉诗勇,王艳伟,杨光参*
摘要:
利用分子梳技术和动态光散射研究了三氯六氨络合钴和DNA分子之间的相互作用. 从荧光显微镜中可直接观察到经YOYO-1染色后?-DNA分子的平均伸直长度比未染色的DNA分子增长约30%. 这是由于荧光染料YOYO-1分子插入双链DNA的碱基对中从而使DNA分子的长度增加. 另外研究了染色后的DNA-[Co(NH3)6]3+混合物的伸直长度随三氯六氨络合钴浓度的变化关系,实验结果表明当钴离子浓度从0增加到3 μmol/L,其混合物的伸直长度由原来的20.9 μm减小到5.9 μm. 利用动态光散射技术证实了这个结果,发现在相同的实验条件下加入钴离子前后DNA-Co(NH3)6]3+混合物的流体力学半径从203.8 nm减小到39.26 nm.
关键词:  分子梳,三氯六氨络合钴,凝聚,动态光散射,荧光显微镜
DOI:10.1088/1674-0068/25/03/359-364
分类号: