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首页> 外文期刊>Journal of Biomolecular Structure and Dynamics >Structural variability of DNA-containing Mg-pyrophosphate microparticles: optimized conditions to produce particles with desired size and morphology
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Structural variability of DNA-containing Mg-pyrophosphate microparticles: optimized conditions to produce particles with desired size and morphology

机译：含DNA的Mg-焦磷酸微粒的结构变异性：优化的条件，以产生具有所需尺寸和形态的颗粒

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摘要

Our previous studies demonstrated the formation of structurally diverse DNA-containing microparticles (DNA MPs) in PCR with Mg-pyrophosphate (MgPPi) as the structure-forming component. These DNA MPs were referred to major structural types: microdisks (2D MPs) with nanometer thickness and 3D MPs with sophisticated morphology and constructed from intersecting disks and their segments. Little is known about factors that influence both the morphology and size of DNA MPs, and the present study was aimed at fulfilling this gap. We showed that the addition of Mn2+ cations to PCR mixtures caused the profound changes in MPs morphology, depending on DNA polymerase used (KlenTaq or Taq). Asymmetric PCR with 20-fold decrease in the concentration of one of two primers facilitated the predominant formation of microdisks with unusual structure. The addition of 1 mM Na-pyrophosphate to PCR mixtures with synthesized DNA and subsequent thermal cycling (10-15 cycles) were optimal to produce microdisks or nanometer 3D particles. Using electron microscopy, we studied also the structure of inorganic micro- and nanoparticles from MgPPi, formed during multiple heating and cooling cycles of a mixture of Mg2+ and Na-pyrophosphate in various regimes. Also, we found the conditions to yield planar (Mg center dot Mn)PPi nanocrystals (diameter similar to 100 nm and thickness similar to 10 nm) which efficiently adsorbed exogenous DNA. These inorganic nanoparticles are promising for DNA delivery in transfection studies. Mechanisms to be involved in structural modifications of MPs and perspectives of their practical application are discussed.
机译：我们之前的研究表明，用Mg-焦磷酸盐（MgPPI）在PCR中形成在结构上不同的DNA微粒（DNA MPS）作为结构成分。这些DNA MPS被称为主要结构类型：微小结构（2D MPS），具有纳米厚度和具有复杂形态的3D MP，并由交叉盘及其段构造。对影响DNA MPS的形态和大小的因素众所周知，目前的研究旨在满足这种差距。我们表明，根据使用的DNA聚合酶（Klentaq或Taq），向PCR混合物添加MN2 +阳离子引起了MPS形态的深刻变化。两种引物中的一个浓度下降的不对称PCR促进了具有异常结构的微小微小的主要形成。将1mM Na-焦磷酸盐与合成的DNA和随后的热循环（10-15个循环）的加入加入1mM-焦磷酸盐以产生微小微小循环或纳米3D颗粒。使用电子显微镜，我们还研究了MgPPI的无机微颗粒和纳米颗粒的结构，在多种加热和冷却循环期间形成的Mg2 +和Na-磷酸盐的冷却循环。此外，我们发现屈服的条件是屈服的平面（Mg中心Mn）PPI纳米晶体（直径与100nm和厚度类似的厚度，类似于10nm），其有效地吸附了外源性DNA。这些无机纳米粒子在转染研究中是有希望的DNA递送。讨论了涉及MPS结构修改的机制以及其实际应用的观点。

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