水牛梭形住肉孢子虫缓殖子抗原的纯化及SDS—PAGE分析

应用葡聚糖凝胶柱层纯化了水牛梭形住内孢子虫缓殖子抗原，并应用ＳＤＳ－ＰＡＧＥ分析了缓殖子可溶性粗抗原及纯化抗原的蛋白组分，结果表明：缓殖子可溶性粗抗原含有１４种蛋白成分，分子量范围为４５．９－７１．２ＫＤ，经过ＳｅｐｈａｄｅｘＧ－１００柱层析纯化扣的抗原在ＳＤＳ－ＰＡＧＥ图谱上显示５条蛋白带，分子量分别为５８ＫＤ，６１ＫＤ，６４ＫＤ，６６ＫＤ，７０ＫＤ。     

从虫体提纯HBsAg基因表达产物的新方法

感染含乙型肝炎病毒表面抗原（ＨＢｓＡｇ）基因的粉纹夜蛾重组核型多角体病毒ＴｎＮＰＶ－Ｈｈｓ８５－ＯＣＣ￣＋的粉纹夜蛾幼虫先经匀浆澄清处理后，再用单克隆抗体亲和柱层析的方法提纯ＨＢｓＡｇ蛋白．提纯的蛋白经ＳＤＳ－ＰＡＧＥ，出现３条电泳带，分子量分别为２４ＫＤ，２７ＫＤ和４５ＫＤ．用此法从虫体提纯ＨａｓＡｇ基因表达产物，具有简单、快速、高效的特点．     

基因工程菌E.Coli2426／pMN表达产物的纯化

用基因工程菌Ｅ．Ｃｏｌｉ２４２６／ｐＭＮ高效表达了麦芽糖结合蛋白—人神经生长因子（β）融合蛋白，菌体经超声波破碎后，上清液经Ａｍｙｌｏｓｅ亲和柱一步即可获得ＳＤＳ－ＰＡＧＥ纯蛋白质，回收率为１０％左右，按标准分子量计，该融合蛋白（５５ＫＤａ）确是麦芽糖结合蛋白（４２ＫＤａ）与人神经生长因子（β）（１３ＫＤ）的络合物     

小麦叶绿体psbA基因的克隆及表达

用ＰＣＲ直接从小麦叶绿体基因组扩增到完整的ｐｓｂＡ基因，并构建了ｐｓｂＡ基因的克隆ｐＴＤ１Ⅱ。为研究ｐｓｂＡ基因在Ｅ·ｃｏｌｉ中的表达，将完整的ｐｓｂＡ基因正向插入高表达载体ｐＫＫ２２３－３中构建表达克隆ｐＫＴＤ１。含有重组表达质粒的转化细胞经ＩＰＴＧ诱导后提取总蛋白，经ＳＤＳ－ＰＡＧＥ分析，小麦叶绿体ｐｓｂＡ基因能在Ｅ·ｃｏｌｉ中表达３２ｋＤ的Ｄ１蛋白，表达的Ｄ１蛋白占细菌总蛋白的３％以上。     

人胰岛素样生长因子—Ⅰ的基因克隆,融合表达及产物纯化

利用聚合酶链式反应（ＰＣＲ）方法,以ＩＧＦ－ＩｃＤＮＡ为模板,扩增ＩＧＦ－Ｉ的基因。在序列测定确证后,将其定向克隆到载体ｐＥｘＳｅｃＩ上,构建了ＩＧＦ－Ｉ融合表达载体ｐＥｘＩＧＦ,经ＩＰＴＧ诱导表达后,ＳＤＳ－ＰＡＧＥ分析表明：含有重组表达质粒的菌株可表达出约３２．５８％的２６ＫＤ融合蛋白,且主要以可溶性形式存在。经ＩｇＧ－Ｓｅｐｈａｒｏｓｅ亲和层析一步纯化后,可获得纯度约９０％的融合蛋白,经Ｗｅｓｔｅｒｎ－Ｂｌｏｔ分析,免疫学活性呈阳性反应。     

金黄色葡萄球菌核酸酶及其突变体酶动力学性质的比较

将金黄色葡萄球菌核酸酶的类似物及其不同点突变体的基因进行表达，经Ｂｉｏ－Ｒｅｘ７０柱层析及尿处理进行分离纯化，在ＳＤＳ－ＰＡＧＥ上显示一条蛋白带，相对分子质量约为１７ｋＤ。测试了它们的比活力，Ｋｍ，Ｖｍａｘ，Ｋｉ等酶动力学参数。结果表明，ＳＮａｓｅＲ与ＳＮａｓｅ的催化活性无显著差异，但ＳＮａｓｅＲ对底物的亲和力明显增加。     

短裙竹荪（Dictyophor a duplicata）多糖的研究（I）：Dd多糖的分离…

竹荪子实体经热水提取，乙醇沉淀，蛋白酶法和Ｓｅｖａｇ法相结合去蛋白，再通过ＤＥＡＥ－Ｓｅｐｈａｄｅｘ Ａ－２５和Ｓｅｐｈａｄｅｘ Ｇ－２００柱层析纯化，得到竹荪多糖（简称Ｄｄ）纯品，经凝胶过滤法和聚丙烯酰胺凝胶电泳法测定表明，Ｄｄ多糖为均一的物质，Ｄｄ经紫外扫描，无核酸和蛋白质的特征吸收峰，ＩＲ－４００扫描表明典型多糖吸收峰，Ｄｄ的相对分子质量约为１９６０００。     

人胰岛素样生长因子—I的基因克隆融合表达及产物纯化

利用聚合酶链式反应（ＰＣＲ）方法,以ＩＧＦ－ＩｃＤＮＡ为模板,扩增ＩＧＦ－Ｉ的基因,在序更测定确证后,将其定向克隆到载体ｐＥｘＳｅｃｌ上,构建了ＩＧＦ－Ｉ融合表达载体ｐＥｘＩＧＦ,经ＩＰＴＧ诱导表达后,ＳＤＳ－ＰＡＧＥ分析表明：含有重组表达质粒的菌株可表达出约３２．５８％的６ＫＤ融合蛋白,且主要以可溶性形式存在。经ＩｇＧ－Ｓｅｐｈａｒｏｓｅ亲和层析一步纯化后,可获得纯度的９０％的融合蛋白,经Ｗｅｓ     

短裙竹荪（Dictyophora duplicata）多糖的研究（Ⅰ）──Dd多糖的分离、纯化及其部分理化性质

竹荪子实体经热水提取，乙醇沉淀，蛋白酶法和Ｓｅｖａｇ法相结合去蛋白，再通过ＤＥＡＥ－ＳｅｐｈａｄｅｘＡ－２５和ＳｅｐｈａｄｅｘＧ－２００柱层析纯化，得到竹荪多糖（简称Ｄｄ）纯品。经凝胶过滤法和聚丙烯酰胺凝胶电泳法测定表明，Ｄｄ多糖为均一的物质。Ｄｄ经紫外扫描，无核酸和蛋白质的特征吸收峰，ＩＲ－４００扫描表明有典型多糖吸收峰，Ｄｄ的相对分子质量约为１９６０００。     

上海四膜虫酚溶性染色质非组蛋白的研究

收集对数生长期的上海四膜虫，分离细胞核，制备染色质并以酚抽提制得酚溶性染以质非组蛋白，经ＳＤＳ－ＰＡＧＥ测得四膜虫酚溶性染色非组蛋白有六条明显的蛋白带，其表观量为８８ＫＤ，７９ＫＤ，７０ＫＤ，５２ＫＤ，３８ＫＤ，２４ＫＤ，与鼠肝酚溶性染色质非组蛋白具有明显的５条的分子量有显著的差异。     

Enhanced expression of H-2K and H-2D antigens on reticulocytes infected with Plasmodium yoelii

The 17XNL strain of Plasmodium yoelii induces a highly effective and permanent T-cell dependent immunity in mice of the CBA strain; the lethal variant P. yoelii 17XL and P. berghei (ANKA) fail to activate an effective immune response in the same host. These differences in immunogenicity are unexplained. We recently observed that in CBA/CaJ mice the intracellular blood stages of P. yoelii 17XNL were almost exclusively within reticulocytes whereas lethal P. yoelii 17XL and P. berghei (ANKA), at comparable stages of infection, were predominantly erythrocytic. Induction of a reticulocytosis converted the normally lethal P. yoelii 17XL infection into a nonlethal one, and reticulocytic P. yoelii was shown to be more immunogenic than the erythrocytic form. Since one of the differences between reticulocytes and erythrocytes that might have influenced the development of immunity was greater expression of MHC antigens of the former cell type we examined the expression of H-2K, H-2D and Ia on reticulocytes infected with P. yoelii 17XNL. These cells showed a very marked increase in H-2K and D antigen expression compared to normal reticulocytes or erythrocytes. No Ia was detected. Red blood cells (RBC) infected with lethal P. yoelii 17XL or P. berghei showed no increase in H-2K or H-2D antigen expression. Finally, the level of expression of H-2K on P. yoelii 17XNL parasitized red blood cells from different strains of mice correlated closely with the ability of these strains to control the infection.     

Malaria parasites--discovery of the early liver form

Infections of mammalian malaria parasites start when sporozoites from an infected anopheline mosquito are injected into the bloodstream of the host. The sporozoites enter the hepatocytes and become transformed into exoerythrocytic schizonts. Since the discovery of the primate parasite Plasmodium cynomolgi in monkey hepatocytes and the rodent parasite Plasmodium berghei in hamster hepatocytes, the ultrastructure of these stages has been extensively studied both in primate and rodent plasmodia. These observations relate only to the development of the exoerythrocytic schizont 25 h after sporozoite injection until the final maturation (of P. berghei) 50 h post-inoculation. Recently, we have studied the route of entry of sporozoites across the cellular lining of liver sinusoids and invasion of the liver parenchymal cells by using transmission electron microscopy. The results of these studies in combination with other physiological experiments strongly suggested that the sporozoite was initially harboured by the Kupffer cell, from which the parasite escaped into the neighbouring hepatocyte. The migration of sporozoites from liver sinusoids to hepatocytes can be achieved within a few minutes. We present here the first ultrastructural observations on the natural transformation of intrahepatocytic sporozoites into exoerythrocytic forms in vivo, using the rodent malaria parasite P. berghei in a laboratory host, the Brown Norway rat. These observations complete the search for the final link in the life cycle of malaria parasites.     

高温敏核不育水稻幼苗生长发育的热激反应

高温敏感核不育水稻培矮 64S 在热激处理后, 幼苗叶蛋白质组分与其可育条件下( 22℃ )相比, 表现出明显差异:在分子量为 97. 4~ 14. 4 KD 范围内共表现出 6 条蛋白质区带的变化, 其中 4 条新增加, 2 条减弱. 在热激条件下, 与常规稻湘晚籼 2号相比较, 培矮 64S 亦表现出了 7条蛋白质区带的变化.这些蛋白质组分的改变必然引起体内某些酶系统的变化,而影响幼苗的正常生长发育, 使幼苗生长受到明显抑制.     

豇豆种子萌发进程中蛋白质组分的时空变化

以豇豆品种(早翠和矮虎)为材料,通过SDS-PAGE技术获得3个时期(干种子期、吸胀期、发芽期)和2个部位[胚(胚芽、胚根和胚轴)和子叶]的3种蛋白质组分(水溶、盐溶和碱溶)的电泳图谱.结果表明:品种间条带分布差异最大的时期是干种子期,部位是胚,组分为盐溶蛋白质.在萌发进程中,相对分子质量为60.4KD、55.4KD和49.7KD的3个条带没有时空变化,推测其是豇豆的保守性蛋白或结构蛋白.组分中以水溶蛋白所显条带数居多,但品种间多态性不丰富,早翠有两个特异带,而矮虎仅一个;盐溶蛋白组分所显品种间条带多态性丰富,早翠有107.8KD、18.3KD、15.7KD、14.0KD、12.8KD和11.4KD6个特异带,矮虎有41.0KD、27.3KD和14.4KD3个.故在豇豆种子萌发进程中,各蛋白质组分在品种内和品种间的时序和空间表达上,既显示出一致性又显示出差异性.由于对干种子期胚或子叶盐溶蛋白质组分的电泳分析,既显示品种遗传稳定性又能反映品种特异性,故均适宜于豇豆基因型判别,但基因型遗传关系分析宜以胚为材料,品种纯度检测宜用子叶为材料,不能两者混合取材,吸胀后的种子蛋白质组分分析不能反映基因型差异.     

Genome sequence and comparative analysis of the model rodent malaria parasite Plasmodium yoelii yoelii

Species of malaria parasite that infect rodents have long been used as models for malaria disease research. Here we report the whole-genome shotgun sequence of one species, Plasmodium yoelii yoelii, and comparative studies with the genome of the human malaria parasite Plasmodium falciparum clone 3D7. A synteny map of 2,212 P. y. yoelii contiguous DNA sequences (contigs) aligned to 14 P. falciparum chromosomes reveals marked conservation of gene synteny within the body of each chromosome. Of about 5,300 P. falciparum genes, more than 3,300 P. y. yoelii orthologues of predominantly metabolic function were identified. Over 800 copies of a variant antigen gene located in subtelomeric regions were found. This is the first genome sequence of a model eukaryotic parasite, and it provides insight into the use of such systems in the modelling of Plasmodium biology and disease.     

盐胁迫下水稻、番茄蛋白质变化的电泳分析

水稻品种 90 1 7经盐胁迫后 ,分子量约为 45 KD蛋白质减少 ,约为 38KD蛋白质增加 ,同时出现了分子量与调渗蛋白相类似的 2 6 KD蛋白质 ,随着盐胁迫浓度的提高及胁迫时间的加长 ,上述蛋白质谱带变化更为明显 .番茄品种白果强丰经盐胁迫后其真叶中分子量约为1 4KD和 1 7KD蛋白质减少 ,同时诱导出一条分子量约为 2 2 KD的新带 .     

盐胁迫下水稻和黑麦幼苗蛋白质组份的变化

耐盐的779水稻幼苗中,盐胁迫后诱导出66kD/P15\5和26kD/p16.0的特异蛋白组份,并有多种蛋白质组份消失和含量减少现象,而在盐敏感的早花2号水稻中,虽然也有少数蛋白质组份被诱导产生,但没有检测到66kD和26kD两种蛋白质组份,并且双向SDS－PAGE谱型相对稳定,一般耐盐的黑麦幼苗中经盐胁迫后也诱导出26kD蛋白组份,因此,66kD,26kD两种蛋白组份可能与耐盐性有关。     

酮替酚和氯喹合并作用对约氏疟原虫抗性株红内期超微结构的研究

酮替酚作用下约氏疟原虫抗性株的超微结构变化与约氏敏感株相似,都表现为有髓鞘状的多层膜结构;而酮替酚和氯喹合并作用下的抗性株有明显不同于单独酮替酚和氯喹作用下的变化:大量类似粗面内质网结构的同心圆排列。     

酮替酚和氯喹合并作用对约氏疟原虫抗性株红内期超微结构的研究

酮替酚作用下约氏疟原虫抗性株的超微结构变化与约氏敏感株相似,都表现为有髓鞘状的多层膜结构;而酮替酚和氯喹合并作用下的抗性株有明显不同于单独酮替酚和氯喹作用下的变化:大量类似粗面内质网结构的同心圆排列。