O_2~-·与红细胞膜阴离子转运蛋白相互关系的研究

利用４,４′二硫氰２,２′二磺基芪（ＤＩＤＳ）对带３（ｂａｎｄ３）蛋白阴离子转运功能的专一抑制作用,研究了超氧阴离子（Ｏ－２·）进入红细胞对带３蛋白的依赖关系;同时根据红细胞在等渗氯化铵溶液中溶血的ｔ１／２,探讨了细胞外大量产生Ｏ－２·对带３蛋白转运阴离子功能的影响。结果表明：４０μｍｏｌ／ＬＤＩＤＳ完全抑制带３蛋白的阴离子转运功能,但并不影响Ｏ－２·进入细胞,证明Ｏ－２·进入细胞对ｂａｎｄ３蛋白无依赖关系;Ｏ－２·通过氧化修饰红细胞膜蛋白的－ＳＨ,使带３蛋白的阴离子转运能力下降,并可用－ＳＨ还原剂ＤＴＴ与巯基乙醇修复。     

人红细胞带3蛋白胞质段的克隆及其表达

人红细胞带3蛋白胞质段(cytoplasmic domain of band 3, cdb3)起着将膜与膜骨架、细胞内环境相联系的重要作用. 以带3蛋白全长基因为模板,用PCR方法扩增出cdb3片段,克隆至pRSET表达质粒上,转化大肠杆菌BL21(DE3). 转化菌经诱导表达出较高含量的cdb3蛋白,纯化后,测得对醛缩酶有抑制活性.     

水稻温敏型突变体叶片间断失绿的超微结构

在短时降温诱导下,水稻温敏型突变体１１０３ｓ（Ｏｒｙｚａ　ｓａｔｉｖａ　ｓｓｐ．ｉｎｄｉｃａ）植株间断失绿性状表达（临界温度２３．１℃）过程中,叶绿体含量的增减与叶色变化相符。电镜观察发现,性状表达时叶片间断失绿区叶绿体内部结构发生退化,呈现基粒垛叠片层数的异常减少,或基粒消失仅剩基粒残迹,有的甚至整个叶绿体为高电子密度的囊泡状结构。但在同一叶片的绿区,叶绿体仅表现基粒片层数减少、排列不规则,嗜锇小球聚集。在叶片失绿区的复绿过程中,叶绿体的这些变化又可逆转,内部结构重建,最后整个叶绿体结构基本恢复正常。水稻温敏突变体１１０３ｓ叶片间断失绿性状表达过程,实质上是一个由温度调控的叶绿体结构退化与修复的可逆过程。     

恙螨染色体分带的初步研究

本文报道应用胰酶法对微红纤恙螨Leptotrombidium rubellum,苍白纤恙螨L. pallidum和小板纤恙满L. scutellarc进行G带带型分析.三种恙螨染色体分别显示17、21、19条深带带纹,用CS-190机对每条显带的染色体进行薄层扫描,结果每一条深带显示一个峰,对微红纤螨和巨螯齿恙螨Odontaearus majestivus进行BSG法的C显带实验,均未见到带纹,从C带结果及对敬红纤慧螨染色体扫描电镜的初步观察结果,提示恙螨染色体可能为泛着丝粒类型,本文根据恙螨染色体的分带情况,探讨了几种恙螨间的亲缘关系以及恙螨染色体的研究在分类上的意义.     

Incorporation of microcrystals by growing protein and virus crystals

In the course of time-lapse video and atomic force microscopy (AFM) investigations of macromolecular crystal growth, we frequently observed the sedimentation of microcrystals and three-dimensional nuclei onto the surfaces of much larger, growing protein or virus crystals. This was followed by the direct incorporation over time of the smaller crystals into the bulk of the larger crystals. In some cases, clear indications were present that upon absorption of the small crystal onto the surface of the larger, there was proper alignment of the respective lattices, and consolidation proceeded without observable defect formation, i.e., the two lattices knitted together without discontinuity. In the case of at least one virus crystal, cubic satellite tobacco mosaic virus (STMV), addition of three-dimensional nuclei and subsequent expansion provided the principal growth mechanism at high supersaturation. This process has not been reported for growth from solution of conventional crystals. In numerous other instances, the lattices of the small and larger crystals were obviously misaligned, and incorporation occurred with the formation of some defect. This phenomenon of small crystals physically embedded in larger crystals could only degrade the overall diffraction and materials properties of macromolecular crystals.     

Comparison of DNA-protein interactions in intact nuclei from avian liver and erythrocytes: A cross-linking study

DNA-protein cross-linkages were formed in intact nuclei of chicken erythrocytes and liver cells by the action of cis-diammine dichloroplatinum (II). Most cross-linked proteins were components of the nuclear matrix, and their heterogeneity reflected the different complexity of liver and erythrocytes matrices, respectively. Some basic proteins, including histones, were also cross-linked, particularly in erythrocyte nuclei. South-Western blotting revealed that a variety of proteins isolated from the cross-linked liver nuclei recognized DNA specifically. In this group of proteins two relatively abundant, acidic, species of 38 and 66 kDa, respectively, might represent novel DNA-binding proteins from the nuclear matrix. In the case of erythrocytes, only the basic proteins showed a DNA-recognition capacity, and among them there were some unidentified species, absent from liver. Lamin B2 was cross-linked but was unable to recognize DNA, and the same was true for other abundant, cross-linked proteins from both types of nuclei. This led to the hypothesis that for some DNA-nuclear matrix interactions the aggregation typical of matrix proteins is essential for the specificity of DNA recognition. Hybridization analysis of the DNA isolated from the cross-linked complexes showed that SARs (scaffold attachment regions) and telomeric sequences were well represented in the cross-linked fragments, that the cross-linked DNA of liver was partially different from that of erythrocytes and that two defined SAR sequences were found to be present only in the cross-linked DNA. These results are in agreement with the present views on DNA-nuclear matrix interactions, which are usually studied on isolated nuclear matrices or purified proteins. Instead, our results provide experimental evidence obtained directly from intact nuclei. © 1996 Wiley-Liss, Inc.     

Liver Nuclear Nadph-Cytochrome P-450 Reductase May be Involved in Redox Cycling of Bleomycin-Fe(III), Oxy Radical Formation and DNA Damage

When NADPH-cytochrome P-450 reductase isolated from rat liver microsomes was aerobically incubated with bleomycin, FeCl₃, NADPH and DNA parallel NADPH and oxygen were consumed and malondialdehyde was formed. A similar parallelism of NADPH- and oxygen-consumption and malondialdehyde formation was observed when ceil nuclei isolated from rat liver were incubated under the same conditions. The formation of malondialdehyde which was identified by HPLC and which was most likely released from oxidative cleavage of deoxyribose of nuclear DNA required oxygen, bleomycin, FeCl₃ and NADPH. This indicates that a nuclear NADPH-enzyme, presumably NADPH-cytochrome P-450 reductase, is able to redox cycle a bleomycin-iron-complex which in the reduced form can activate oxygen to a DNA-damaging reactive species. The data suggest that the activity of this enzyme in the cell nucleus could play an important role in the cytotoxicity of bleomycin in tumor cells.     

Comprehensive maturity of nuclear pore complexes regulates zygotic genome activation

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西双版纳楝径向生长和木质部发生季节动态及其对环境因子的响应

热带森林在全球碳循环过程中起着重要的作用，研究热带树木的生长动态及其对环境因子的响应，有助于预测全球变化下热带森林固碳能力的变化。目前对热带树木径向生长以及木质部发生季节动态的研究相对较少。利用微树芯和生长环法对西双版纳热带植物园内的落叶树种楝(Melia azedarach)2021年的年内径向生长和木质部发生季节动态进行监测，结合环境因子的监测，分析了楝的径向生长过程的驱动因子。结果表明：2021年楝的扩大细胞在3月1日(年积日(DOY):60.2±1.6)开始出现，细胞壁加厚在12月11日(DOY:345.4±3.1)结束，年生长量为7.76 mm,最大生长速率为0.039 mm/d。生长环监测显示，楝在3月18日(DOY:76.6±8.6)左右开始生长，10月13日(DOY:286.4±6.8)生长结束，年径向生长量为(6.25±2.35)mm,最大生长速率为(0.056±0.02)mm/d。楝的径向生长速率与降水(r=0.77,P<0.001)、平均温度(r=0.61,P<0.05)和最低温度(r=0.67,P<0.001)均呈显著正相关，与风速(r=-0....