电子显微学报
電子顯微學報
전자현미학보
JOURNAL OF CHINESE ELECTRON MICROSCOPY SOCIETY
2014年
4期
330-336
,共7页
郑露露%江俊龙%柯震栋%楼江潭%何俊杰%王艳伟
鄭露露%江俊龍%柯震棟%樓江潭%何俊傑%王豔偉
정로로%강준룡%가진동%루강담%하준걸%왕염위
介电常数%电泳迁移率%DNA凝聚%精胺
介電常數%電泳遷移率%DNA凝聚%精胺
개전상수%전영천이솔%DNA응취%정알
dielectric constant%electrophoretic mobility%DNA condensation%spermine
DNA在四价平衡离子精胺溶液中,会发生电荷逆转,这个现象无法用经典的Poisson-Boltzman理论解释,一般认为与离子-离子关联、介电环境等有关。当改变溶液的介电常数,DNA-精胺复合物呈现不同的逆转及凝聚状态。本文首先利用动态光散射研究不同介电环境(溶液中加入乙醇、甘氨酸及6-氨基己酸)下DNA-精胺的电泳迁移率的变化,发现乙醇会有利于DNA发生电荷逆转,在0?5 mmol·L-1的精胺浓度下,DNA的电泳迁移率为-0?6×10-4 cm2 V-1 S-1。当加入90%的乙醇后,DNA的电泳迁移率变为0?11×10-4 cm2 V-1 S-1,然而向精胺-DNA溶液中加入甘氨酸,6-氨基己酸等两性物质之后, DNA的电泳迁移率随着两性物质浓度的增加而向负向移动。在此基础上,作者用原子力显微镜观察精胺-DNA复合物在不同浓度乙醇及两性物质下的表面形貌变化,发现随着乙醇浓度的增加,精胺-DNA复合物的凝聚形态会更加紧凑;而随着两性离子浓度的增加,精胺-DNA复合物的凝聚形态变得越来越松散,说明介电环境在精胺导致DNA构象变化及电荷逆转中起着一定的作用。研究不同介电环境下DNA的电泳迁移率及凝聚构象的影响,能对认识DNA凝聚的动力学特性提供帮助,而且有助于获得理想的非病毒载体,促进基因治疗的发展。
DNA在四價平衡離子精胺溶液中,會髮生電荷逆轉,這箇現象無法用經典的Poisson-Boltzman理論解釋,一般認為與離子-離子關聯、介電環境等有關。噹改變溶液的介電常數,DNA-精胺複閤物呈現不同的逆轉及凝聚狀態。本文首先利用動態光散射研究不同介電環境(溶液中加入乙醇、甘氨痠及6-氨基己痠)下DNA-精胺的電泳遷移率的變化,髮現乙醇會有利于DNA髮生電荷逆轉,在0?5 mmol·L-1的精胺濃度下,DNA的電泳遷移率為-0?6×10-4 cm2 V-1 S-1。噹加入90%的乙醇後,DNA的電泳遷移率變為0?11×10-4 cm2 V-1 S-1,然而嚮精胺-DNA溶液中加入甘氨痠,6-氨基己痠等兩性物質之後, DNA的電泳遷移率隨著兩性物質濃度的增加而嚮負嚮移動。在此基礎上,作者用原子力顯微鏡觀察精胺-DNA複閤物在不同濃度乙醇及兩性物質下的錶麵形貌變化,髮現隨著乙醇濃度的增加,精胺-DNA複閤物的凝聚形態會更加緊湊;而隨著兩性離子濃度的增加,精胺-DNA複閤物的凝聚形態變得越來越鬆散,說明介電環境在精胺導緻DNA構象變化及電荷逆轉中起著一定的作用。研究不同介電環境下DNA的電泳遷移率及凝聚構象的影響,能對認識DNA凝聚的動力學特性提供幫助,而且有助于穫得理想的非病毒載體,促進基因治療的髮展。
DNA재사개평형리자정알용액중,회발생전하역전,저개현상무법용경전적Poisson-Boltzman이론해석,일반인위여리자-리자관련、개전배경등유관。당개변용액적개전상수,DNA-정알복합물정현불동적역전급응취상태。본문수선이용동태광산사연구불동개전배경(용액중가입을순、감안산급6-안기기산)하DNA-정알적전영천이솔적변화,발현을순회유리우DNA발생전하역전,재0?5 mmol·L-1적정알농도하,DNA적전영천이솔위-0?6×10-4 cm2 V-1 S-1。당가입90%적을순후,DNA적전영천이솔변위0?11×10-4 cm2 V-1 S-1,연이향정알-DNA용액중가입감안산,6-안기기산등량성물질지후, DNA적전영천이솔수착량성물질농도적증가이향부향이동。재차기출상,작자용원자력현미경관찰정알-DNA복합물재불동농도을순급량성물질하적표면형모변화,발현수착을순농도적증가,정알-DNA복합물적응취형태회경가긴주;이수착량성리자농도적증가,정알-DNA복합물적응취형태변득월래월송산,설명개전배경재정알도치DNA구상변화급전하역전중기착일정적작용。연구불동개전배경하DNA적전영천이솔급응취구상적영향,능대인식DNA응취적동역학특성제공방조,이차유조우획득이상적비병독재체,촉진기인치료적발전。
If DNA are in solution including tetravalent counter-ions spermine, the inversion of DNA charge will happen. This phenomenon cannot be explained by classical Poisson-Boltzman theories. Generally considered associated with the interaction of ion-ion, dielectric environment and so on. When the dielectric constant of solution change, DNA-spermine compounds present different reverse and condensed state. In this paper, first of all we study the change of the electrophoretic mobility of DNA-spermine on different dielectric environment (adding ethanol solution, glycine and 6-amino caproicacid) by using dynamic light scattering. We found that ethanol are good for the charge reversal of spermine induced DNA. When the concentration of spermine is 0?5 mmol·L-1 , DNA electrophoresis mobility is -0?6 × 10 -4 cm2 V-1 S-1 , when we added 90% ethanol to the DNA solution, DNA electrophoresis mobility is 0?11 × 10 -4 cm2 V-1 S-1 . However, when we added zwitterionic species to spermine-DNA solution ( glycine, amino caproicacid) , DNA electrophoresis mobility will decrease with increasing the concentration of zwitterionic species. On this basis, we observed the morphologies of spermine - DNA complexes under different concentrations of ethanol and zwitterionic species by atomic force microscopy. We found that the condensed morphologies of spermine-DNA complexes will be more compact with ethanol concentration increasing. And the condensed morphologies of spermine - DNA complexes become more loose with zwitterionic species concentration increasing. The results show that the dielectric environment plays a certain role on DNA conformation change and charge reverse. We study the electrophoresis mobility and condensed conformation of DNA under different dielectric environment. This can help to recognize the dynamic characteristics of DNA condensation and obtain ideal non viral vector to promote the development of gene therapy.