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首页> 外文期刊>Nucleic Acids Research >CROSSLINKING OF PROGESTERONE RECEPTOR TO DNA USING TUNEABLE NANOSECOND, PICOSECOND AND FEMTOSECOND UV LASER PULSES
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CROSSLINKING OF PROGESTERONE RECEPTOR TO DNA USING TUNEABLE NANOSECOND, PICOSECOND AND FEMTOSECOND UV LASER PULSES

机译：使用可调节的纳秒级，皮秒级和飞秒级紫外激光脉冲使孕酮受体与DNA交联

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摘要

UV laser crosslinking is a potentially powerful tool to investigate transient DNA-protein interactions and binding kinetics in intact cells. As the processes underlying UV laser crosslinking are not fully understood, we have performed a study of the influence of laser pulses with different physical parameters on crosslinking of the progesterone receptor to an oligonucleotide containing a hormone-responsive element. We also studied the influence of the various parameters on the amount of laser-irradiated DNA that can be correctly primer extended as an operational measurement of DNA integrity. A strong influence of pulse intensity and pulse length on the crosslink yield was found, likely due to a change in the 'two photon' processes responsible for crosslinking. The highest efficiency of protein crosslinking to DNA was achieved with femtosecond pulses and should be sufficient to enable use of this technique for in vivo studies.
机译：UV激光交联是研究完整细胞中瞬时DNA-蛋白质相互作用和结合动力学的潜在强大工具。由于尚未完全了解UV激光交联的基本过程，因此我们进行了研究，研究了具有不同物理参数的激光脉冲对孕酮受体交联到含有激素反应性元件的寡核苷酸的影响。我们还研究了各种参数对激光辐照DNA量的影响，可以正确引物延伸以作为DNA完整性的操作度量。发现脉冲强度和脉冲长度对交联产率的强烈影响，可能是由于负责交联的“两个光子”过程发生了变化。通过飞秒脉冲达到了蛋白质与DNA交联的最高效率，应该足以使该技术用于体内研究。

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