激光育种机理非线性研究

本文对激光与ＤＮＡ分子相互作用,考虑了随机力作用,建立了ＤＮＡ分子在激光作用下的Ｆｏｋｋｅｒ－ｐｌａｎｃｋ方程（ＦＰＥ）。分析了ＤＮＡ分子系统的ＦＰＥ势函数,对激光育种中的ＤＮＡ遗传变异的随机不确定性现象进行了解释。     

激光育种机理的动力学研究

对ＤＮＡ分子系统的动力学研究表明,ＤＮＡ分子系统存在双稳势。激光与ＤＮＡ作用,可激发ＤＮＡ分子系统进入混沌状态,从而引起遗传变异。若无激光作用,生物体系是一个稳定体系。     

半导体激光辐照DNA的光谱分析

本文通过半导体激光等辐照源对ＤＮＡ辐照,研究激光、紫外、普通红光对ＤＮＡ吸收光谱的影响。发现激光（６６０ｎｍ）与紫外（峰值波长２５４ｎｍ）均能使ＤＮＡ吸收峰值改变,表明它们均能被ＤＮＡ吸收,与ＤＮＡ发生作用,从而使ＤＮＡ构型发生变化,但激光、紫外与ＤＮＡ作用机理是不同的。而普通红光（峰值波长６６０ｎｍ）对ＤＮＡ光谱无甚影响,这说明了激光与ＤＮＡ作用的非线性共振吸收存在。并发现在激光辐照ＤＮＡ时,激光剂量不同以及ＤＮＡ溶液浓度不同,对ＤＮＡ光谱的影响也不同。这些结果对激光育种和激光生物学实验有一定参考价值。     

激光作用DNA的非线性量子遗传突变理论分析

采用Ｗｉｇｎｅｒ表示和Ｈｕｓｉｍｉ表示分析了ＤＮＡ量子系统，并给出了在激光作用下，不同自由度的ＤＮＡ分子的经典和量子系统非线性共振的突变条件。这一研究结果有助于激光对ＤＮＡ的损伤、对生物遗传致突变性诱变以及致癌效应的解释。     

激光与DNA作用方程的稳定态分析

本文在庞小峰改进后的Ｙｏｍｏｓａ模型基础上,引进了激光与ＤＮＡ分子系统作用的非线性方程。     

激光与DNA作用系统非线性共振实验初探

用紫外可见分光光度计,对受Ｈｅ－Ｎｅ激光辐照过的ＤＮＡ溶液进行扫描。发现２０９ｎｍ吸收峰降低了１０％,表明６３２．８ｎｍＨｅ－Ｎｅ激光波ＤＮＡ吸收而引起其构象变化。Ｈｅ－Ｎｅ激光波长６３２．８ｎｍ,约为２０９ｎｍ的３倍,恰好为我们理论研究的激光与ＤＮＡ相互作用非线性系统的超谐波共振波长。因此该实验结果使我们的理论结果得到了初步验证。     

二自由度DNA系统受激光激励动力学研究

本文将DNA分子作二自由度振子系统近似,建立了激光与DNA相互作用系统的动力学方程。并进行了主共振,超谐波共振及组合共振的分析,从而进一步解释了激光育种中的频率现象。     

DNA芯片技术

ＤＮＡ芯片技术利用固定化在固体表面的高密度ＤＮＡ分子微点阵来进行生化分析。从生物体中提取的样品ＤＮＡ或ＲＮＡ作为靶分子,荧光或其它方法标记后,与微点阵上互补的探刈杂交,从而产生异源双链。因为在微点阵上,某一特定位置年的核苷酸序列是已知的,所以通过对微点阵每一位点的荧光强度进行检测,便可能样品进行定性及定量分析,检测技术包括激光共聚焦扫描和电耦合设备（ＣＣＤ）成像等等。ＤＮＡ芯片根据控针成分的不同大     

原子力显微镜研究不同温度下质粒DNA结构变化

采用原子力显微镜（ＡＦＭ）在无水乙醇中成象了ｐＢＲ３２２质粒ＤＮＡ。在不同温度范围加热云母表面的ＤＮＡ溶液,并在相应温度下干燥使ＤＮＡ充分展开地固定在云母基底上。ＡＦＭ观察结果表明：在３０～４０℃,质粒ＤＮＡ主要是超螺旋和环状分子;在４０～５０℃,环状分子开环形成线形分子;　在７０～８０℃,这些线形分子全部变性,解链形成无规线团。我们通过测定ｐＢＲ３２２质粒ＤＮＡ的熔融曲线,发现了两个突变点,对应于ＡＦＭ所观察到的质粒ＤＮＡ在加热过程中由环状变成线形、双螺旋解开成单链的两个结构变化过程。这是首次通过ＡＦＭ直接观察到质粒ＤＮＡ在不同温度下的结构变化。     

激光辐照对离体质粒DNA的断裂效应

利用低功率的Ｈｅ－ Ｎｅ 激光辐照离体质粒ＤＮＡ,并用离子束和紫外线为对照,分析了质粒ＤＮＡ的单双链断裂效应。结果表明：Ｈｅ－ Ｎｅ 激光辐照可诱发质粒ＤＮＡ 断裂,且断裂频率随辐照剂量增大而提高;Ｈｅ－ Ｎｅ 激光辐照后的质粒超螺旋双链ＤＮＡ存活率剂量曲线不同于紫外线和氮离子束。Ｈｅ－ Ｎｅ 激光诱发双链断裂的频率低于氮离子束和紫外线。     

高频激光-DNA分子相互作用体系相空间的局域特征

利用激光与DNA分子相互作用的动力学模型,借助适当的数值方法,讨论了系统动力学演化及相空间特性。结果表明：高频激光可使DNA分子的运动状态发生变化;特别地,相空间分析表明,高频激光的作用会使DNA分子运动在一些特定状态之间转变。从而可说明高频激光作用下DNA分子呈现新的有序运动现象。     

Study of the orientation of DNA molecules in an electrical field by anisotropy of electrical conductivity of their aqueous solution

Electro-conductive anisotropy of DNA solution caused by the molecular orientation in the external electric field was investigated. The dependence of a relative change of the conductivity of aqueous-salt DNA solution on the electric field in the amplitude range 0-700 V/cm and in the frequency range 100 Hz-10 kHz was studied. It was pointed out that the field thermal effect is an overcoming factor when the orientation of DNA molecules is investigated by field-free relaxation.     

Quantifying Double-Strand Breaks and Clustered Damages in DNA by Single-Molecule Laser Fluorescence Sizing

Fluorescence from a single DNA molecule passing through a laser beam is proportional to the size (contour length) of the molecule, and molecules of different sizes can be counted with equal efficiencies. Single-molecule fluorescence can thus determine the average length of the molecules in a sample and hence the frequency of double-strand breaks induced by various treatments. Ionizing radiation-induced frank double-strand breaks can thus be quantified by single-molecule sizing. Moreover, multiple classes of clustered damages involving damaged bases and abasic sites, alone or in combination with frank single-strand breaks, can be quantified by converting them to double-strand breaks by chemical or enzymatic treatments. For a given size range of DNA molecules, single-molecule sizing is as or more sensitive than gel electrophoresis, and requires several orders-of-magnitude less DNA to determine damage levels.     

Ice-water interface migration by temperature controlling for stretching of DNA molecules

This report shows a new DNA stretching method using migration of an ice-water interface. DNA molecules were stretched accompanying the migration of the solid-liquid interface and immobilized in frozen area. This simple method needs no chemical modification to keep DNA in the stretched form. For full stretching of DNA molecules, one terminus of the DNA molecules were anchored on silanized substrate. The anchored DNA molecules were stretched by freezing the DNA solution. The stretched DNA molecules were observed after sublimation of the frozen solution keeping its stretched form on silanized surface which had no attractive interaction with DNA molecules except for the SH-modified terminus in solution. An infrared (IR) laser beam was introduced to a frozen DNA solution through an objective lens for local area melting of the solution. Scanning of the laser irradiation caused stretching and enclosing of DNA molecules in the frozen area followed by migration of the solid-liquid interface.     

DNA analysis by microfabricated capillary electrophoresis device.

The LIGA (Lithographie Galvanoformung Abformung) process using synchrotron radiation lithography is applied to the microfabrication of capillary array electrophoresis (CAE) device. Laser-induced fluorescence detection system for the CAE device has been constructed by the modification of laser confocal fluorescence microscopy. DNA molecules were detected during migrating in the microchannels filled with polymer separation matrices under electric field to optimize the separation conditions for DNA analysis. Based on this observation, we demonstrated that microfabricated CAE device is realized the fast separation of DNA.     

Conformational correlatives of DNA band compression and bidirectional migration during field inversion gel electrophoresis, detected by quantitative video epifluorescence microscopy.

Individual DNA molecules in the Mb size range were monitored by epifluorescence video microscopy during field inversion gel electrophoresis (FIGE). DNA migrating in an agarose gel gives rise to characteristic V-conformational elements and when doing so exhibits a reduced mobility. When the V-conformational elements per DNA molecule are few, the degree of retardation appears proportional to the number of V's, and since larger DNA species exhibit more V's, to DNA size. For a particular pulse frequency, the proportionality breaks down progressively as the number of V-conformational elements per DNA molecule increases. The loss of proportionality between DNA length and migration rate is being correlated with the macroscopically observed loss of electrophoretic size discrimination known as band compression. For a particular pulsing frequency and size class of DNA, the loss of size discrimination is thought to be due to the different orientations of migration, caused by the asymmetric distribution of V-conformational elements when the number of these elements is moderate. Small and very large DNA by contrast migrate with the direction of the biased field. These events, analyzed by microscopic measurement, are consistent with the known macroscopically observed double-valued mobilities in FIGE.     

Direct laser trapping of single DNA molecules in the globular state.

A sharply focused laser is able to trap small particles at the laser focal point due to the difference in refractive index of the particles and that of the surrounding medium. This technique, called laser trapping, can be used to manipulate animal or bacterial cells without any contact and has been widely applied in biological research. However, it has been difficult to trap biological macromolecules such as DNA molecules, because these molecules give a low difference in refractive index and cannot overcome Brownian motion. DNA molecules can be transformed to a condensed globular state. This transformation results in a higher refractive index of DNA due to its increased density. We demonstrate in this paper that a single DNA molecule can be optically trapped using a Nd:YAG laser (1064 nm wavelength) upon transformation from the coiled state to the globular state.     

Theory of acoustic mode vibrations of DNA fibers

A previous model for acoustic mode vibrations of a DNA molecule in water is extended to the case of an array of many DNA molecules, as occurs in the fibers studied in most experimental work on DNA. The acoustic modes of this system are found to consist of coupled modes of water sound vibrations and DNA acoustic modes. This model is used to study the electrostatic coupling of acoustic vibrations to the relaxational modes of the orientational degrees of freedom of the water molecules. It is found that the long-range or macroscopic electric field generated by the acoustic mode vibrations of the water-DNA system gives too small a damping and frequency shift of the acoustic modes to account for the observations on DNA fibers. Therefore, the observed damping and frequency shifts are most likely due to either friction between the surrounding water and the vibrating DNA, or coupling to the water orientation degrees of freedom resulting from the short range (i.e., screened) Coulomb interaction. The latter explanation (which is most likely the correct one) implies that the relaxation time of the hydration shell water is longer than the observed relaxation time by a factor of the static dielectric constant of the hydration water.     

Electrodeless dielectrophoresis of single- and double-stranded DNA

Dielectrophoretic trapping of molecules is typically carried out using metal electrodes to provide high field gradients. In this paper we demonstrate dielectrophoretic trapping using insulating constrictions at far lower frequencies than are feasible with metallic trapping structures because of water electrolysis. We demonstrate that electrodeless dielectrophoresis (EDEP) can be used for concentration and patterning of both single-strand and double-strand DNA. A possible mechanism for DNA polarization in ionic solution is discussed based on the frequency, viscosity, and field dependence of the observed trapping force.     

Effect of pulsed and reversing electric fields on the orientation of linear and supercoiled DNA molecules in agarose gels

When linear or supercoiled DNA molecules are imbedded in agarose gels and subjected to electric fields, they become oriented in the gel matrix and give rise to an electric birefringence signal. The sign of the birefringence is negative, indicating that the DNA molecules are oriented parallel to the electric field lines. If the DNA molecules are larger than about 1.5 kilobase pairs, a delay is observed before the birefringence signal appears. This time lag, which is roughly independent of DNA molecular weight, decreases with increasing electric field strength. The field-free decay of the birefringence is much slower for the DNA molecules imbedded in agarose gels than observed in free solution, indicating that orientation in the gel is accompanied by stretching. Both linear and supercoiled molecules become stretched, although the apparent change in conformation is much less pronounced for supercoiled molecules. When the electric field is rapidly reversed in polarity, very little change in the birefringence signal is observed for linear or supercoiled DNAs if the equilibrium orientation (i.e., birefringence) had been reached before field reversal. Apparently, completely stretched, oriented DNA molecules are able to reverse their direction of migration with little or no loss of orientation. If the steady-state birefringence had not been reached before the field reversal, complicated orientation patterns are observed after field reversal. Very large, partially stretched DNA molecules exhibit a rapid decrease in orientation at field reversal. The rate of decrease of the birefringence signal in the reversing field is faster than the field-free decay of the birefringence and is approximately equal to the rate of orientation in the field (after the lag period).(ABSTRACT TRUNCATED AT 250 WORDS)