若干种枝角类卵鞍表面亚显微结构的比较研究

以扫描电镜对发头裸腹溞、蚤状溞、隆线溞指名亚种以及隆线溞东湖一亚种的卵鞍分别进行了比较研究,发现卵鞍表面的亚显微结构有明显的差异,但亚种之间差异较小。隆线溞指名亚种与隆线溞东湖一亚种二者的卵鞍在光学显微镜下看不出有什么差异,然而亚显微结构却互不相同。这就为东湖一新亚种的确定提供了一项可靠的依据,同时也进一步证实卵鞍的超微结构确可作为枝角类分类的表征。     

人肌肌酸激酶胍变性时的失活与构象变化的比较研究

应用二阶导数光谱、紫外差吸收光谱和荧光光谱等监测手段,研究了人肌肌酸激酶在盐酸胍溶液中的构象变化。二阶导数光谱结果表明,若以6M盐酸胍中肌酸激酶酪氨酸残基的暴露程度为100％,则天然酶酪氨酸残基的暴露程度只有2％。而紫外差吸收光谱和荧光光谱的变化与兔肌肌酸激酶的结果相似。比较不同胍浓度下人肌肌酸激酶的失活与构象变化,表明酶的失活先于构象变化。同时还测定了不同浓度胍溶液中人肌酶的失活与构象变化的速度常数。结果表明以几种方法测定的构象变化均为单相的一级过程,而酶的失活却呈现了由快慢两相组成的一级反应过程。比较同浓度胍溶液中的失活速度与构象变化速度,发现酶失活的快相反应速度常数比构象变化的速度常数大1—2个数量级,慢相速度常数与构象变化速度常数相近。上述结果进一步支持了酶的活性部位构象柔性的观点。     

人α1/158Ⅴ型(α1c型)干扰素基因在大肠杆菌中的表达及其产物的初步纯化

采用DNA聚合酶链反应(PCR)技术,将本实验室从中国人胎肝细胞染色体DNA中发现和分离的IFN-α1/158V基因的原始克隆,改造成适于进行非融合蛋白原核表达的结构形式,并在大肠杆菌中获得高效表达。测得重组IFN-α1/158V的抗病毒活性为1.9×10~7单位/升菌液。随后又采用以单克隆抗体亲和层析为主的纯化流程对表达产物进行初步纯化,获得了在SDS-聚丙烯酰胺凝胶电泳上呈现单一条带的纯化产物。     

利用带有原核增强子样序列的新型载体在大肠杆菌中高效表达人α肿瘤坏死因子

将去除信号肽的人肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)cDNA插入到带有原核增强子样序列Px的新型表达载体pBV320中,使TNF cDNA 5′端直接置于大肠杆菌trp启动子下游,采用37℃恒温培养,使TNF在大肠杆菌中获得了高效表达,表达活性达1.35(±0.17)×10~6U/L菌液。表达的TNF-α对L929细胞的毒性作用可被抗人肿瘤坏死因子-α的单克隆抗体所中和。表达菌裂解液作SDS-聚丙烯酰胺凝胶电泳,显示有一条分子量与TNF分子量吻合、约为17000道尔顿的蛋白带。利用DEAE-Sepharose阴离子交换层析及Sephacryl S-200凝胶过滤对上述重组人TNF-α进行纯化,获得电泳纯产品,比活性为1.48×10~6U/mg。     

室旁核在剌激猫肾神经传入纤维引起的血压效应中的应用

The purpose of this study is to investigate the role of paraventricular nucleus of the hypothalamus (PVN) and alpha 1 adrenergic receptor of PVN in the pressor responses to stimulation of renal afferent nerve in alpha 1-chloralose-anesthetized cats with carotid sinoaortic denervation and vagotomy. The pressor response to stimulation of renal afferent nerve consisted of a primary and a second components. The primary component response was completely blocked while the second component was not blocked by autonomic blocking agents (hexomethonium and atropine). Bilateral lesions of PVN greatly attenuated the pressor response before and after autonomic blockade. Intracerebroventricular and PVN injection alpha 1, adrenergic antagonist (prazosin) significantly decreased in the pressor response to stimulation of renal afferent nerve. These results indicate that paraventricular nucleus of the hypothalamus and alpha 1 adrenergic receptors in central nervous system, especially in PVN, play an important role in the pressor responses to stimulation of renal afferent nerve.     

大鼠中枢神经系统衰老的形态学研究Ⅳ.大脑皮质锥体细胞树突棘的数量变化

用6、12与31个月的雄性Wistar大鼠的大脑Krieg 2、3区皮质,对其V层大锥体细胞的五段50μm长度内的树突棘做形态学定量研究。在Golgi法的切片中共计数了三个年龄组的151个细胞的725段树突的棘密度。结果表明,老年大鼠比成年和青年大鼠的棘密度普遍下降。其中以基树突与侧树突棘度下降最显著(减少24％左右),顶树突只中段有明显减少。老年大鼠锥体细胞还常出现胞体、树突及其分支的明显形态改变。     

Effect of disulfide cross-linking between alpha and delta subunits on the properties of the F1 adenosine triphosphatase of Escherichia coli

Under very mild oxidizing conditions the delta subunit of the F1-ATPase of Escherichia coli can be crosslinked by a disulfide linkage to one of the alpha subunits of the enzyme. The cross-linked ATPase resembles the native enzyme in the following properties: specific activity; activation by lauryldimethylamine N-oxide (LDAO); binding of aurovertin D and ADP; cross-linking products with 3,3'-dithiobis(succinimidyl propionate); binding to ATPase-stripped everted membrane vesicles and the N,N'-dicyclohexylcarbodiimide sensitivity of the rebound enzyme. However, the rebound crosslinked ATPase differed from the native enzyme in lacking the ability to restore NADH oxidation - and ATP hydrolysis-dependent quenching of the fluorescence of quinacrine to ATPase-stripped membrane vesicles. It is proposed that the delta subunit is involved in the proton pathway of the ATPase, and that this pathway is affected in the alpha delta-cross-linked enzyme. The mechanism for activation of the ATPase by LDAO was examined. Evidence against the proposal of L?tscher, H.-R., De Jong, C. and Capaldi, R.A. (Biochemistry (1984) 23, 4140-4143) that activation involves displacement of the epsilon subunit from an active site on a beta subunit was obtained.     

单纯疱疹病毒Ⅱ型DNA克隆片段的限制酶切分析

本文采用10种限制性内切核酸酶分析了单纯疱疹病毒Ⅱ型(HSV-2)335株DNA BglⅡ8种克隆片段共12个重组质粒。发现了HSV-2 DNA L链末端区域有限制酶切位点的变异并对HSV-2 DNA单一序列区域和末端重复区域的稳定性进行了初步讨论。本文应用末端标记技术和末端杂交分析法对编码糖蛋白D的HSV-2 DNA BglⅡL片段和含有形态学转化基因的HSV-2DNA BglⅡN片段进行了详细的限制性内切核酸酶物理图谱分析。