CAJ | 학술논문

【目的】在生理状态下利用激光镊子拉曼光谱系统对单个酿酒酵母孢子萌发过程进行实时监测,探讨掩盖在群体平均信息下的个体生命信息。【方法】将葡萄糖溶液加入酿酒酵母孢子悬液中诱导孢子萌发,在孢子萌发过程中每隔30 min取样并利用激光镊子拉曼光谱系统测定单个酵母孢子的拉曼光谱。【结果】单细胞实时平均拉曼光谱可显示孢子萌发过程中细胞内生物分子的变化：萌发期内分别归属于 DNA、蛋白质的722 cm-1,1006 cm-1峰的峰高基本不变,随后在生长期上升明显,说明生长期胞内开始大量复制DNA,并合成蛋白质；归属脂类的1751 cm-1峰的减弱趋势明显,可能是胞内脂类物质消耗造成的。整个萌发生长过程中,源自葡糖糖和海藻糖的858 cm-1,908 cm-1,1084 cm-1,1118 cm-1等峰强度先下降后上升,表明在适宜的生长条件下,海藻糖可能转化为单糖被细胞吸收利用,当营养物质逐渐被消耗时,细胞会再次累积海藻糖以抵抗外界不利条件。【结论】激光镊子拉曼光谱技术可反映胞内生物大分子的活动规律,获知单个酵母孢子萌发过程中物质变化的丰富信息,是探索单个活细胞实时生化变化的有效工具。
【목적】재생리상태하이용격광섭자랍만광보계통대단개양주효모포자맹발과정진행실시감측,탐토엄개재군체평균신식하적개체생명신식。【방법】장포도당용액가입양주효모포자현액중유도포자맹발,재포자맹발과정중매격30 min취양병이용격광섭자랍만광보계통측정단개효모포자적랍만광보。【결과】단세포실시평균랍만광보가현시포자맹발과정중세포내생물분자적변화：맹발기내분별귀속우 DNA、단백질적722 cm-1,1006 cm-1봉적봉고기본불변,수후재생장기상승명현,설명생장기포내개시대량복제DNA,병합성단백질；귀속지류적1751 cm-1봉적감약추세명현,가능시포내지류물질소모조성적。정개맹발생장과정중,원자포당당화해조당적858 cm-1,908 cm-1,1084 cm-1,1118 cm-1등봉강도선하강후상승,표명재괄의적생장조건하,해조당가능전화위단당피세포흡수이용,당영양물질축점피소모시,세포회재차루적해조당이저항외계불리조건。【결론】격광섭자랍만광보기술가반영포내생물대분자적활동규률,획지단개효모포자맹발과정중물질변화적봉부신식,시탐색단개활세포실시생화변화적유효공구。
Objective]The germination process of Saccharomyces cerivisiae spores under physio-logical conditions is monitored real-time using laser tweezers Raman spectroscopy to obtain in-formation on metabolic activity of individual spores which could be masked by bulk measure-ment.[Methods]The germination was initiated by addition of glucose solution to spore suspen-sion and samples were harvested at a 30 min interval during germination process.Raman spec-tra of individual spores at different time points were recorded.[Results]The Raman spectra of the single spore reflected the change in bio-molecule content during the process of spore germination:The intensities of band at 722 cm-1 and 1006 cm-1 (assigned to DNA and protein,re-spectively)did not change significantly during germination period,whereas they increased dra-matically during the outgrowth period.This in-dicated the DNA replication and protein biosyn-thesis occurred during outgrowth period.The in-tensities of 1751 cm-1 band (associated with lipids)decreased substantially,which might be as the result of consumption of lipids.During the whole germination and outgrowth process,the intensities of bands at 858 cm-1 ,908 cm-1 ,1084 cm-1 and 1118 cm-1 originated from glucose and trehalose decreased at first and then iecreased.This phenomenon suggested that under the favorable growing conditions,trehalose may be converted into monosaccharide and then be uti-lized.When nutritional materials are gradually consumed,the cells would accumulate trehalose again in order to resist adverse condition.[Conclusion]The experimental results show that the LTRS technique can reveal the metabolic activity of bio-molecules and provides rich informa-tion on the change in compound content within single spores during the germination process. Therefore,the LTRS can serve as a valuable tool for the real time monitoring of dynamic chan-ges in bio-molecules at single cell level.