鱼类染色体的荧光显带研究

应用GC碱基特异性荧光染料色霉素A,辅以AT减基特异性荧光染料Hoechst33258,DAPI或喹吖因对鲤鱼,鲫鱼,大鳞副泥鳅和的有丝分裂染色体及黄鳝的有丝分裂和减数分裂染色体进行了荧光显带研究,结果发现,色霉素A3可以特异性地显示鱼类有丝分裂及减数分裂各个时期核仁组织区NORS的存在,Hoechst33258,DAPI或喹吖因则使这些区域（NORs)淡染,大鳞副泥鳞的染色体NORs 分布位置具有性别,根据实验结果,对有关鱼类染色体的荧光染色研究及其应用进行了讨论。     

一种野外制备染色体的新方法-骨髓整体低渗法

本文以整体低渗的方法进行哺乳与鸟类的染色体制备,以探讨一种在野外条件下,无法进行离心时,适合细胞遗传学研究的染色体制备新方法。应用该方法可在野外考察中,同时开展细胞遗传学方面的工作在某些特殊研究课题中,该方法有着广阔的应用前景。     

黑斑蛙骨髓细胞染色体标本制备的新方法

染色体标本制备是黑斑蛙分子细胞遗传学研究的基础。为了简化操作程序,缩短实验周期,建立了黑斑蛙染色体制备的一种新方法——骨髓细胞体外短时培养与秋水仙素同步处理法。该方法操作简便,重复性较好,可在较短时间里制备出分裂相较多且形态良好的蛙染色体标本,适用于核型分析、染色体荧光原位杂交、染色体显微分离和单染色体文库构建等多个方面的研究。     

草鱼染色体的包装（间期—中期）

以草鱼ＺＣ７９０１细胞株为材料,观察鱼类细胞从间期染色质到中期染色体的包装过程。主要通过（１）分裂期与间期细胞融合,诱导染色体早熟凝集;（２）染色体“伸长”处理;（３）培养细胞的低渗处理;（４）染色质辅展等方法,制作染色体标本,进行扫描和透射电镜观察。观察表明,鱼类染色质的基本结构与哺乳类细胞相同,也是直径约１０ｎｍ的核丝。染色体的色装有两种形式：一种是多级螺旋化形成直径约３００ｎｍ的染色单体,     

一种从鱼类卵巢制备地高辛标记mtDNA探针的简易方法

一种从鱼类卵巢制备地高辛标记ｍｔＤＮＡ探针的简易方法＊ＡＳＩＭＰＬＥＭＥＴＨＯＤＦＯＲＧＥＴＴＩＮＧＤＩＧＯＸＩＧＥＮＩＮＬＡＢＥＬＩＮＧＰＲＯＢＬＥＯＦｍｔＤＮＡＦＲＯＭＯＶＡＲＹＯＦＦＩＳＨ关键词鱼类,线粒体ＤＮＡ,地高辛标记ＫｅｙｗｏｒｄｓＦｉ...     

一种改良的鱼类染色体富集制片方法

运用淋巴细胞分离液处理黄鳝肾细胞悬浮液后,成功地分离了其中红细胞和淋巴细胞,以富集的淋巴细胞进行染色体制片,可避免有核红细胞的影响,获得高分裂指数的黄鳝染色体标本,该方法简便,结果稳定,亦适用于其他鱼类,两栖类和鸟类的染色体制片。     

制备青虾染色体方法新探

1方法1.1秋水仙素注射挑选活动力强、性成熟的青虾,每个体重(5±0.2)g,确定秋水仙素的注射剂量为0.1mL,注射质量分数分别是1×10-3mg/g、2×10-3mg/g、3×10-3mg/g4×10-3mg/g、5×10-3mg/g、6×10-3mg/g(体重),用微量注射器于第1与第2腹足之间腹肌注射,对照组注射0.1mL的生理盐水,作用时间24h,取出卵巢、精巢、触角腺、肝胰腺、心脏、鳃,分别制片。1.2细胞低渗处理除去表面脂肪,用生理盐水洗涤后,转移到尖底离心管里,加入适量的生理盐水,超声震荡仪打散细胞,在进行1500r/min的离心8min。然后用吸管小心吸去上清液。加入0.4%KCl至8mL,于37…     

淡水涡虫染色体的制备方法

本试验证明,采用空气干燥法和压片法,利用淡水涡虫的再生组织制备染色体是可行的,能够得到较好的效果。     

鱼类染色体G—显带的Brd U—BSG方法及白鲢G—带模式图的初步建立

本文报道了一种显示鱼类染色体G-带的BrdU-BsG方法。采用肾细胞短期培养,收获前12小时加入BrdU,使终浓度为10μg/ml。制片经HCl、Ba(OH)_2处理,4×SSC温育。Giemsa染色,显示出白鲢的G-带。其带纹细致清晰,一个细胞的单倍染色体上显示带纹达200条以上,是目前已报道的鱼类多重带中带纹最多的,且反差明显,带纹有特征性,结果较稳定。根据实验结果初步建立了白鲢的G-带模式图。     

Acettc-Orcein: A New Stain-Fixative for Chromosomes

A new stain-fixative method for chromosomes, namely acetic-orcein, is described, which gives results that are equally good in fresh and permanent preparations. A 45% acetic and 1% orcein content is recommended as a standard solution. For salivary glands of Drosopkila a 2% stain gives the best results, and with the two species D. melanogaster and D. miranda the acetic strength has been raised to 70% with advantage. The addition of chloroform proves necessary for hardening in species of Sciara. Acetic-orcein is equally good for rapid chromosome counts. For root tips the addition of 1 cc. of N HC1 solution to 10 cc. of the standard solution together with gentle heating of the tissues in a drop of the mixture assists in the softening and separation of cells necessary for chromosome study. Orcein can also be used successfully in other combinations such as acetic-propionic or acetic-lactic. The latter is useful for making preparations that do not require ringing. Preparations so made keep from 7 to 14 days.     

用家蝇取代果蝇制作唾腺多线染色体新技术

组建学生科技兴趣小组,对家蝇唾液腺染色体制片方法进行研究,开发了制片新技术,即加热处理家蝇三龄幼虫,然后用改进的缓冲液进行解离前预处理,再酸性解离、染色、观察、永久制片、显微照相.利用新方法,学生均能一次就获得细胞膜破裂、背景清晰、染色体分散良好、横纹清楚的制片.     

一种检测人中期染色体原位切口移位的新方法

本研究首次详细描述了在BrdU替代4个细胞周期以上的中期染色体上进行原位染色体切口移位的方法。研究证明,切口移位效率达峰值的最适温度为15—20℃,最佳时间为10—15分钟,DNasc Ⅰ的最佳浓度为2ng/ml。用本方法进行的人外周血淋巴细胞染色体的原位切口移位带型表明,原位切口移位的染色体带型特征与已知的G带、R带有较明显的差异,是另一种新的带型。本方法较用’H-dTTP或Bio-dUTP作标记物进行染色体原位切口移位更简便、快速,不仅可用于活性基因在不同种类基因组内的分布研究,而且可能在细胞遗传学和分子生物学研究中具有更广泛的用途。     

Preparation of Insect Chromosomes for Immunolabeling

We describe a method for isolating chromosomes from testes of the desert locust, Schistocerca gregaria, and their subsequent incubation with antibodies directed against chromosomal proteins. The procedure involves hypotonic pretreatment of the germ cells, centrifugation onto coverslips in a cytocentrifuge and immunolabeling, while still unfixed, using a chromatin-stabilizing buffer. In the present case, an antibody specific for the acetylated isoforms of his tone H4 was tested. After the antibody treatment, the preparations are fixed using formaldehyde, stained with a DNA-specific fluorescent dye and mounted. Analysis of the preparations revealed good preservation of chromosome structure in prophase spermatogonia and late prophase I spermatocytes. Fully condensed chromosomes were not observed and are probably lost during preparation. The bright fluorescence of the autosomes indicates that the reaction between the antibody against acetylated histone H4 and its chromosomal antigen is not impeded. In contrast, the X univalent remained unstained with the exception of a small terminal band. Thus, cytospin preparations of locust germ cells allow high resolution immunolabeling with antibodies against chromosome-associated proteins.     

A New Method for Preparation of Undecalcified Bone Sections

A new method for preparation of sections of undecalcified bone is described. Samples of ovine bone were embedded in methylmethacrylate and thick-sectioned with a cutoff machine or commercial band saw. Composite slides were prepared by gluing white acrylic to glass using cyanoacrylate glue. Bone sections were glued to the composite slide and then surface polished by grinding or ultramilling. The polished surface of the section was then etched and stained. The techniques described in this paper reduce the time spent grinding or milling sections and improve resolution of surface-stained features of undecalcified bone sections.     

一个新的粗糙脉孢霉基因组DNA制备方法

粗糙脉孢霉基因组DNA的制备多数费工费时,并且需要液氮研磨。而一般实验室没有液氮设备。本文提供了一个不需要液氮研磨就可以快速制备粗糙脉孢霉基因组DNA的方法,使用裂解液、石英砂和苯酚振荡裂解细胞壁,然后用苯酚／氯仿抽提DNA。利用PCR从基因组扩增出粗糙脉孢霉Ⅰ号染色体右臂上的校孔转运蛋白基因片段。