Pre-translational induction of pentoxyresorufin O-depentylase by pyridine.

Pentoxyresorufin O-depentylase activity, mainly associated with phenobarbital-inducible cytochrome P450IIB1 (designated CYP2B1), was increased after a single treatment of pyridine (250 mg/kg, i.p.), and further increased by repeated treatments for 5 days. The catalytic activity and immunoreactive protein of CYP2B recognized by polyclonal antibodies were significantly induced by a relatively high dose of pyridine (250 mg/kg, i.p.) while ethanol-inducible cytochrome P450IIE1 (CYP2E1) could be induced by a low dosage (25 mg/kg, i.p.). Unlike CYP2E1 induction without changing its mRNA level, the induction of CYP2B by pyridine was accompanied by an elevation of its mRNA, indicating a pre-translational activation of this enzyme. These results indicate that pyridine induces various isozymes of cytochromes P450 by different induction mechanisms.     

厚朴类有效成分的含量测定及高效液相色谱图

用高效液相色谱法对我国木兰科植物“厚朴类”中的酚类和季胺碱类作了定性和定量分析,并提供色谱图,结果表明不同产地的“厚朴”,图谱相同,仅含量有差别,但不同种的“厚朴”,色谱图不同.此结果可作为评价“厚朴类“药材质量和鉴别的方法.根据各类厚朴有效成分的存在,有一定的规律,为木兰科植物化学分类提供了有价值的资料。     

猪肠毒素源性大肠杆菌K88ac粘附抗原的核苷酸序列分析

 本文对国内流行的猪肠毒素源性大肠杆菌K88ac抗原的结构基因的核苷酸序列进行了测定。该基因由849对核苷酸组成,编码了283个氨基酸的蛋白亚单位及21个氨基酸的信号肽。与国外报道的K88ac序列的不同是我们发现一个碱基的点突变,导致在抗原决定簇内一个氨基酸的改变。从核苷酸序列推导出的氨基酸序列与另两种亚型进行了比较。     

ALLOZYME DIVERSITY AND GENETIC IDENTITY IN SCHIEDEA AND ALSINIDENDRON (CARYOPHYLLACEAE: ALSINOIDEAE) IN THE HAWAIIAN ISLANDS

Genetic diversity of allozymes, genetic identity based on allozyme variability, and phylogenetic relationships were studied with respect to breeding system diversity, population size, and island age in 20 of the 29 species of Schiedea and Alsinidendron (Caryophyllaceae: Alsinoideae), a monophyletic lineage endemic to the Hawaiian Islands. Average levels of genetic variability in Schiedea and Alsinidendron were comparable to or higher than those found in other Hawaiian lineages for which equivalent data are available [Bidens, Tetramolopium, and the silversword alliance (Asteraceae: Madiinae)] and similar to average values for species of dicots. Allozyme variability was strongly dependent on breeding system, which varies widely in the Hawaiian Alsinoideae. Species with autogamous breeding systems showed very low variability, measured as the number of alleles per locus, percent polymorphic loci, and mean heterozygosity per locus. Outcrossing hermaphroditic and dimorphic species (those with gynodioecious, subdioecious, and dioecious breeding systems) showed significantly higher genetic variability. Small population size was associated with lower values for all measures of genetic variability. Nearly half of the species occurring in small populations are also autogamous; thus, both factors may have influenced levels of genetic variability in these species. Founder effect was apparent in one species (Schiedea adamantis), which occurs in a single large population, has a gynodioecious breeding system but a very low genetic variability. Island age appeared to have little effect on genetic variability. Slightly lower values of genetic variability for species occurring on Kaua'i and O'ahu result primarily from the occurrence of autogamous Alsinidendron species on those islands. Values for Nei's genetic identity for different species pairs were 0.201–0.942, a far greater range than in Bidens, the silversword alliance, and Tetramolopium. Using UPGMA clustering, there was only moderate support for relationships detected through cladistic analysis. Nei's unbiased genetic identity (I) was greatest among species with outcrossing breeding systems, which for the most part clustered together. Nei's genetic identities for self-fertilizing species were low, indicating that these species are less similar to one another and to outcrossing species, regardless of their affinities based on cladistic analysis. Parsimony analysis of allele frequency data supported two clades also found in phylogenetic analyses using morphological and molecular data. Clades recognized in parsimony analysis of allele frequencies were those lineages containing selfing species, indicating that conditions favoring fixation of alleles occurred in ancestral species. In contrast, maintenance of high genetic diversity in outcrossing species interferes with recognition of phylogenetic relationships using allozyme variability.     

Helper virus-free transfer of herpes simplex virus type 1 plasmid vectors into neural cells.

Herpes simplex virus type 1 (HSV-1) plasmid vectors have promise for genetic intervention in the brain, but several problems caused by the helper virus have compromised their utility. To develop a helper virus-free packaging system for these vectors, the DNA cleavage/packaging signals were deleted from a set of cosmids that represents the HSV-1 genome. Following cotransfection into cells, this modified cosmid set supported replication and packaging of vector DNA. However, in the absence of the DNA cleavage/packaging signals, the HSV-1 genome was not packaged, and consequently vector stocks were free of detectable helper virus. In the absence of helper virus, the vectors efficiently infected rat neural cells in culture or in the brain with minimal cytopathic effects. beta-galactosidase-positive cells were observed for at least 1 month in vivo, and vector DNA persisted for this period. This system may facilitate studies on neuronal physiology and potential therapeutic applications.     

水稻幼芽细胞生物膜上的赤霉素结合蛋白的结合特性

在水稻 (Oryza sativa)幼芽中存在膜结合的赤霉素结合蛋白 ,其与 GA3 结合的平衡解离常数(Kd)为 6.5× 1 0 -8mol/ L,总浓度为 0 .3 pmol· mg-1 蛋白质。结合蛋白与 GA3 结合活力在 0℃时比 2 5℃时高 1 4 0 %。它与 GA3 结合的最适 p H为 5。 GA3 与此结合蛋白的结合量随反应时间延长而增加 ,1 h达最大值 ,以后又逐渐下降。 IAA、ABA可与 GA3 竞争赤霉素结合蛋白。     

大豆结瘤突变体成分的初步分析及其对硝酸还原酶活性和结瘤的影响

超结瘤大豆（Ｇｌｙｃｉｎｅ ｍ ａｘ （Ｌ．） Ｍｅｒｒ．） ｎｔｓ ３８２ 和不结瘤大豆Ｎｏｄ ４９ 的叶和根组织水提取物经Ｓｅｐｈａｄｅｘ Ｇ２５ 过滤、洗脱,再根据洗脱物对硝酸还原酶（ＮＲ）活性的影响可划分为４ 个组分（ｆｒａｃｔｉｏｎ）样品,即ｎｔｓ ３８２（Ｎｏｄ ４９） Ｆ１、ｎｔｓ ３８２（Ｎｏｄ ４９） Ｆ２、ｎｔｓ ３８２（Ｎｏｄ ４９） Ｆ３ 和ｎｔｓ ３８２（Ｎｏｄ ４９） Ｆ４。其中, ｎｔｓ３８２ Ｆ２ 和Ｆ４ 抑制ＮＲ 活性作用在接种ＵＳＤＡ１１０ 后明显下降, 但接种的ｎｔｓ ３８２ Ｆ２ 却能提高大豆Ｂｒａｇｇ 的结瘤数达一倍, 而接种的ｎｔｓ ３８２ Ｆ３ 和Ｆ４ 的作用不明显。ＮＲ 活性抑制因子不是刺激结瘤的因子, 刺激结瘤的因子主要分布在接种的ｎｔｓ３８２ Ｆ２ 部分中。与这一现象相反, Ｎｏｄ ４９ Ｆ２ 和Ｆ４ 抑制ＮＲ活性的作用在接种后更强, 且也抑制大豆ｎｔｓ ３８２ 的结瘤, 其中Ｎｏｄ ４９ Ｆ４ 抑制结瘤的作用基本不能逆转。抑制结瘤因子主要分布在接过种的Ｎｏｄ ４９ Ｆ４ 部分中     

细胞分裂素生物合成基因在调节一组烟草病理相关蛋白基因表达中的作用

对一组病理相关蛋白基因在烟草 ( N icotiana tabacum cv. Wisconsin 38)中的表达情况进行了研究 ,包括 :碱性几丁质酶、β- 1 ,3-葡萄糖苷酶、渗透蛋白及伸展蛋白。RNA杂交实验表明在正常烟草植株中上述 4个基因具有发育和器官专一性的表达。在含有细胞分裂素生物合成基因的转基因烟草丛生芽中 ,这 4个基因的表达受过量合成的内源细胞分裂素和载体效应的共同调节 ,细胞分裂素降低这些基因的表达 ,而载体效应则促进它们的表达。热激处理也明显降低这 4种基因的表达水平。上述结果表明这些病理相关蛋白基因具有复杂的调控系统     

Optimal conditions for levan formation by an overexpressed recombinant levansucrase

Summary A genetically modified levansucrase, which contained His-affinity tag in its C-terminal, was constructed by PCR reaction using two synthetic primers. This modified protein was produced up to 30 % in total cell protein of E. coli, and was purified by a one-step affinity chromatography. The optimum pH for levan production was pH 5 and the optimum temperature was 0 °C. The higher velocity of levan formation within shorter enzyme reaction times was achieved by increasing the levels of enzyme concentration. The optimal sucrose concentration for levan production was around 20 %. Under these conditions, more than 50 g levan/l was produced.