不同地理单元尖吻蝮蛇毒蛋白组分比较研究

目的探讨不同地理单元尖吻蝮蛇毒蛋白组分有无差异性。方法采用非变性聚丙烯酰胺凝胶电泳(NativePAGE)方法,比较采自安徽黄山(黄山单元)和贵州梵净山(西部单元)尖吻蝮蛇毒组分。结果梵净山尖吻蝮的蛇毒蛋白表达量和条带数目均高于黄山尖吻蝮。结论黄山单元和西部单元的尖吻蝮蛇毒蛋白组分具有差异性。     

尖吻蝮蛇毒研究进展

蛇毒是从毒蛇头部毒腺分泌的有毒液体,是一种重要的天然动物毒素,其主要成分是蛋白质,具有广泛的生物学活性,现已被应用于溶栓、抗癌、止血、止痛等方面。传统上蛇毒分为神经毒、血循毒与混合毒。尖吻蝮是我国十大剧毒蛇之一,其毒液主要作用于血液系统,引起出血、肿胀与局部组织坏死。已经发现尖吻蝮蛇毒多属丝氨酸蛋白酶、金属蛋白酶和磷脂酶屯家族,部分蛋白质可能属于去整合素家族[1]。现就尖吻蝮蛇毒生理活性及药理应用等方面的研究进展综述如下。     

湖南尖吻蝮蛇毒出血毒素的圆二色性和化学修饰

用远紫外ＣＤ谱研究了湖南产尖吻蝮蛇毒的两个出血毒素的溶液构象,计算得ＤａＨＴ－１的α螺旋、β折叠、无规卷曲分别为２３．４％、３１．３％、４５．３％。随ＰＨ的增大或减小,峰位蓝移,酸性条件下的变化比碱性条件下的变化大。构象单元含量计算表明：α螺旋减少,无规卷曲增多,β折叠基本未变,温度和ＰＨ对ＣＤ谱的影响相似,５０℃时峰位蓝移,α螺旋减少,无规卷曲增多,ＥＤＴＡ对ＣＤ谱影响显著,０．０２ｍｏｌ／ＬＥ     

湖南尖吻蝮蛇毒两个出血毒素的纯化和理化性质

经ＳｅｐｈａｄｅｘＧ－７５凝胶过滤,ＱＡＥ－ＳｅｐｈａｄｅｘＡ－５０和ＣＭ－Ｓｅｐｈａｄｅｘ Ｃ－２５离子交换层析的步骤,从湖南产尖吻蝮蛇毒中纯化出两个出血毒素。ＳＤＳ－ＰＡＧＥ测得分子量均为２３．５ｋＤ,ＩＥＦ－ＰＡＧＥ测得等电点分别为５．６和５．２,两者具有相似的氨基酸组成,其中酸性氨基酸分别占２３％和２４％。ＤａＨＴ－１和ＤａＨＴ－２的最小出血剂量（ＭＨＤ）分别为０．５μｇ和０．８μｇ。都具     

尖吻蝮（Deinagkistrodon acutus）生长过程不同时期排毒量及蛇毒组分的变化

对人工培育的4个年龄段尖吻蝮蛇毒和野生成体尖吻蝮蛇毒进行了聚丙烯酰胺凝胶电泳分析。结果显示：3龄以前尖吻蝮蛇毒蛋白电泳图谱,无论是在分带、泳动率及相应组分的量等方面,均呈现一定的差异,但随着尖吻蝮蛇龄的增长,蛇毒蛋白电泳图谱与野生成蛇蛇毒蛋白电泳图谱越来越接近。3龄尖吻蝮生长发育达到性成熟,其蛇毒蛋白电泳图谱与野生成蛇趋于一致。蛇毒组分的变化,提示了尖吻蝮的生长发育进程。     

细菌蛋白的聚丙烯酰胺凝胶电泳鉴定

细菌蛋白的聚丙烯酰胺凝胶电泳鉴定华西医科大学口腔医学院口腔生物医学工程重点实验室成都６１００４１黄毅早在１９０８年,电泳现象就已被发现,Ｔｉｓｅｌｉｕｓ于１９３７年对电泳仪器作了重大改进,随后电泳技术才日益普及并成为鉴定生物大分子的基本工具。Ｆｏｗｌ...     

皖南尖吻蝮蛇毒出血毒素Ⅳ的纯化与初步鉴定

从皖南尖吻蝮蛇（Ａｇｋｉｓｔｒｏｄｏｎａｃｕｔｕｓ）毒液中经ＤＥＡＥ－Ｓｅｐｈａｒｏｓｅ和ＳｅｐｈａｃｒｙｌＳ－２００两步凝胶柱层析首次纯化出一种中分子量出血毒素（简称ＡａＨⅣ）．经ＳＤＳ－ＰＡＧＥ和等电聚焦凝胶电泳测定其分子量为４４ｋＤ,等电点为ｐＨ５．０．从５００ｍｇ粗毒中可获得２０ｍｇＡａＨⅣ纯品．ＡａＨⅣ有较强的出血活性,最小出血剂量（ＭＨＤ）为０．４μｇ．     

皖南尖吻蝮蛇毒出血毒素IV的纯化与初步鉴定

从皖南尖吻蝮蛇毒液中经ＤＥＡＥ－Ｓｅｐｈａｒｏｓｅ和Ｓｅｐｈａｃｒｙｌ Ｓ－２００两步凝胶柱层析首次纯化出一种中分子量血毒素（简称ＡａＨⅣ）。经ＳＤＳ－ＰＡＧＥ和等电聚焦凝胶电泳测定其分子量为４４ｋＤ,等电点为ｐＨ５．０。从５００ｍｇ粗毒中可获得２０ｍｇ ＡａＨⅣ纯品。ＡａＨⅣ有较强的出血活性,最小出血剂量（ＭＨＤ）为０．４μｇ。     

皖南尖吻蝮蛇蛇毒中纤溶组分的荧光特性

用荧光光谱方法研究了皖南尖吻蝮蛇蛇毒中纤溶组分。研究了Ａｃｒ对ＦＰ内源发光的影响,结果表明,每个ＦＰ分子含有多个Ｔｒｐ残基,这些Ｔｒｐ残基约有８３％可被Ａｃｒ所淬灭,而且这些Ｔｒｐ残基位于ＦＰ分子的较疏水环境中。     

尖吻蝮(Deinagkistron acutus)生长过程不同时期排毒量及蛇毒组分的变化

对人工培育的4个年龄段尖吻蝮蛇毒和野生成体尖吻蝮蛇毒进行了聚丙烯酰胺凝胶电泳分析.结果显示3龄以前尖吻蝮蛇毒蛋白电泳图谱,无论是在分带、泳动率及相应组分的量等方面,均呈现一定的差异,但随着尖吻蝮蛇龄的增长,蛇毒蛋白电泳图谱与野生成蛇蛇毒蛋白电泳图谱越来越接近.3龄尖吻蝮生长发育达到性成熟,其蛇毒蛋白电泳图谱与野生成蛇趋于一致.蛇毒组分的变化,提示了尖吻蝮的生长发育进程.     

Polyacrylamide Gel Electrophoresis of Plasteins

A peptic hydrolysate of soybean protein was filtered with Sephadex G–25 and was separated approximately into four fractions (I, II, II, and IV in the order of mol. wt.). Fraction II (av. mol. wt: 1043) and III (av. mol. Wt.: 685) were more plastein-productive than others. When plastein produced from Fraction II with Nagarse was investigated by plate electrophoresis using 7.5% polyacrylamide-gel, the upper limit of the molecular weight was found to be about 25,000. A similar result was obtained also with Fraction III. The increase of molecular weight in the course of the plastein formation with the mixture (substrate) of Fractions II and III was shown that the final product lay mainly in a position between cytochrome c (mol. wt.: 11,700) and Nagarse (mol wt.: 27,600). In addition, the gel-electrophoretic experiments revealed that the most favorable condition for the plastein synthesis were pH 6.5 and 35% in substrate concentration.     

常规聚丙烯酰胺凝胶电泳快速检测胰蛋白酶抑制剂的方法

ＳＤＳ—ＰＡＧＥ－　明胶电泳是检测蛋白酶抑制剂的常用方法,基本原理为：聚丙烯酰胺凝胶中加入ＳＤＳ和明胶,　　ＳＤＳ使蛋白酶抑制剂发生可逆变性。电泳完毕,用　Ｔｒｉｔｏｎ　Ｘ－　１００恢复凝胶板中蛋白酶抑制剂的活性,凝胶板置于蛋白酶溶液里保温,含蛋白酶抑制剂部位的明胶不被蛋白酶水解而呈现蛋白染色带。ＳＤＳ－ＰＡＧＥ一明胶电泳检测蛋白酶抑制剂时,在同一块凝胶板上可同时检测酸性、碱性及中性的蛋白酶抑制剂,但电泳后处理（包括复性,酶解,漂洗和染色）时间长（约２０ｈ）,而且对于同一样品中分子量相近的蛋白酶…     

聚丙烯酰胺凝胶电泳的快速脱色方法

以牛血清白蛋白为材料进行聚丙烯酰胺凝胶电泳(PAGE)和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE),凝胶固定后,用考马斯亮蓝R-250染色后比较传统脱色液(冰乙酸-甲醇溶液)和不同盐溶液(NACL、KCL、CUCL2)的脱色效果的结果表明:PAGE和SDS-PAGE胶,0.25和0.5MOL·L-1NACL,在70℃(PAGE)、50℃(SDS-PAGE)下脱色,约2￣4H,效果好,灵敏度高,背景低。     

High Resolution Polyacrylamide Gradient Gel Electrophoresis

A method was developed for high resolution electrophoresis of proteins in linear gradient (3 to 30%) polyacrylamide gel rods in a neutral phosphate buffer containing 0.1% sodium dodecyl sulfate. Well-defined protein zones were observed and improved resolution was attained especially for low molecular weight proteins in preparations containing a variety of polypeptides, e. g. viruses that are often separated by continuous gel methods. Electropherograms of continuous (8%)and gradient (3 to 30%) gels were made of purified vesicular stomatitis virus, variola virus, Rickettsia rickett-sii, and alpha and beta chains of hemoglobin in order to demonstrate the resolution of the gradient system.     

Separation of Colicins by Polyacrylamide Gel Electrophoresis

S ummary : Colicins can be separated by polyacrylamide gel electrophoresis, so allowing multiple colicinogeny to be detected with a universal indicator strain.     

Detection of Avocado Sunblotch Viroid in Flower Buds by Polyacrylamide Gel Electrophoresis

A Rapid PAGE index method for avocado sunblotch disease for field trees which takes only 6 h, compared with 2—5 days for other previously described tediniques, has been developed and tested. Flower bud? are used instead of leaves as source material. All 52 flower Duu Siimplcs irom known bunblotcn-infcctcd trees tested positive whilst only 37 out of 72 in-fccted leaf samples gave positive results.     

Flat Gel Polyacrylamide Electrophoresis of Porcine Mycoplasmas

The flat gel acrylamide electrophoresis technique was standardized and applied to the comparison of four species of porcine mycoplasmas. Clear differences were observed between these species, and differences were seen among the strains of Mycoplasma hyosynoviae. The clarity of the patterns and the number of bands developed was influenced by the amount of protein in the extract and the age of the culture. The technique allows the comparison of several protein extracts in parallel without the problems associated with the rearrangement of separate gel columns.     

人脑胶质细胞瘤核染色质非组蛋白的聚丙烯酰胺凝胶电泳分析

用不连续SDS-聚丙烯酰胺凝胶电泳分析了人脑胶质细胞瘤与正常脑细胞核NHCP电泳图谱。从二者的NHCP电泳结果表明,脑胶质细胞瘤增加了一条表观分子量为3万的蛋白区带;表观分子量为1.75万～6.5万的蛋白区带染色明显加深。染色质紫外吸收光谱也有明显差异。总之,脑胶质细胞瘤核NHCP发生质与量的变化。     

蛋白质复合体非变性凝胶电泳技术及其应用新进展

非变性凝胶电泳技术是研究蛋白质复合体的强有力工具。重点介绍了蓝绿温和非变性凝胶电泳(BN-PAGE)技术的原理和特点,比较了由BN-PAGE衍生的蓝色温和琼脂糖凝胶电泳、清澈温和非变性凝胶电泳(CN-PAGE)和高分辨清澈温和非变性凝胶电泳(HrCN-PAGE)技术的差异和适用范围,并概括地介绍了这些技术在植物蛋白质复合体研究中应用的新进展。