长吻鮠精子超微结构的观察

用透射电镜和扫描电镜观察了长吻（ＬｅｉｏｃａｓｓｉｓｌｏｎｇｉｒｏｓｔｒｉｓＧｕｎｔｈｅｒ）精子的超微结构。长吻精子具有椭圆的头部和复杂的中片;近侧中心粒和远侧中心粒靠近核的中央;鞭毛呈９＋２轴丝结构,并具有由外膜折迭形成的波浪形的鳍状结构。研究表明,长吻精子具有某些区别于其它鱼类精子的不同结构。     

棘腹硅体内寄生原生动物两新种记述：动鞭毛纲：动基体目：锥体 科;粘孢子…

本文记述棘腹蛙Ｒａｎａ ｂｏｕｌｅｎｇｅｒｉ Ｇｕｎｔｈｅｒ体内寄生的两种文献中未报道过的原生动物,一种是利川锥虫新种,Ｔｒｙｐａｎｏｓｏｍａ ｌｉｃｈｕａｎｅｎｓｉｓ ｓｐ．ｎｏｖ．,其主要特征是,鞭毛较粗,一般有伸出体外,故不形成游离的鞭毛;另一种是蛙两极虫新种,Ｍｙｘｉｄｉｕｍ ｂｏｕｌｅｎｇｅｒｉ ｓｐ．ｎｏｖ．,孢子卵形,两端钝圆,壳厚,壳面光滑,缝线直而宽。     

注射促黄体素释放激素类似物和地欧酮诱导大鳍■和长吻■排卵的研究

对于性成熟的大鳍Ｍｙｓｔｕｓｍａｃｒｏｐｔｅｒｕｓ（Ｂｌｅｅｋｅｒ）野生鱼,单独注射多巴胺的抑制剂地欧酮（ＤＯＭ）不能影响血清促性腺激素（ＧＴＨ）水平,也不能诱导排卵;单独注射类似物ＬＨＲＨ－Ａ,虽能使血清ＧＴＨ水平显著升高,但仅产生较低的排卵率;而当ＤＯＭ与ＬＨＲＨ－Ａ结合注射却显著增强ＬＨＲＨ－Ａ促进血清ＧＴＨ水平升高的作用,并诱导出较高的排卵率。对性成熟的长吻ＬｅｉｏｃａｓｉｓｌｏｎｇｉｒｏｓｔｒｉｓＧｕｎｔｈｅｒ野生鱼,使用ＬＨＲＨ－Ａ＋ＤＯＭ作２次注射诱导排卵的效果也与注射ＬＨＲＨ－Ａ加脑垂体这一传统诱导排卵方法相似。Ｌｉｎｐｅ方法（ＬＨＲＨ－Ａ＋ＤＯＭ,作１次或２次注射）避免了采集、保存脑垂体不便给生产带来的麻烦,在科鱼类的人工繁殖中,具有较高的推广价值。     

长吻Wei幼鱼的赖氨酸需要量

本文通过以添加结晶氨基酸的半精制饲料饲喂长吻Ｗｅｉ（ＬｅｉｏｃａｓｓｉｓｌｏｇｎｉｒｏｓｔｒｉｓＧｕｎｔｈｅｒ）幼鱼的生长试验来确定春赖氨基酸需要量,试验饲料由鱼粉,玉米蛋白,糊精,明胶等组成,通过添加游离氨基酸达饲料中分别含赖氨酸国１．６４,２．０４,２．４４,２．８４和３．２４％五个水平,结果得出：长吻Ｗｅｉ幼鱼最大生长时对饲料中的赖氨酸的需要量为２．５７％,占饲料蛋白的６．５４％。     

几丁质酶和壳聚糖酶对部分乙酰化壳聚糖作用方式的比较

通过对几丁质酶和壳聚糖酶降解部分乙酰化壳聚糖的作用方式的比较,得到几丁质酶切断壳聚糖的ＧｌｃＮＡｃ－ ＧｌｃＮＡｃ 和ＧｌｃＮＡｃ－ ＧｌｃＮ 或ＧｌｃＮ－ ＧｌｃＮＡｃ 糖苷键,而壳聚糖酶切断壳聚糖的ＧｌｃＮ－ ＧｌｃＮ 和ＧｌｃＮ－ ＧｌｃＮＡｃ 或ＧｌｃＮＡｃ－ ＧｌｃＮ 糖苷键,为得到较高聚合度的壳寡糖提供理论基础。     

G蛋白Rap1活化新机制：GEFs研究进展

小分子Ｇ蛋白Ｒａｓ超家庭成员都是通过与鸟苷酸交换因子（ｇｕａｎｉｎｅ ｎｕｃｌｅｏｔｉｄｅ ｅｘｃｈａｎｇｅ ｆａｃｔｏｒｓ,ＧＥＦｓ）结合而被活化,ＧＥＦｓ能增加这类小分子Ｇ蛋白对ＧＴＰ的摄取,并使之形成有活性的ＧＴＰ结合构象。Ｒａｐ１是Ｒａｓ样的小分子Ｇ蛋白。迄今为止,已发现Ｇ３Ｇ、ＣａｌＤＡＧ－ＧＥＦ１、Ｅｐａｃ、ｃＡＭ－ＧＥＦ１、ｃＡＭＰ－ＧＥＦⅡ、ｎＲａｐＧＥＰ、ＧＦＲ可特异的活化Ｒａｐ     

蛋白聚糖研究进展

在糖复合物（ｇｌｙｃｏｃｏｎｊｕｇａｔｅ）中,蛋白聚糖（ｐｒｏｔｅｏｇｌｙｃａｎ,ＰＧ）与糖蛋白的不同是其核心蛋白上共价连接有糖胺聚糖（ｇｌｙ－ｃｏｓａｍｉｎｏ－ｇｌｙｃａｎ,ＧＡＧ）链,数量可由１条至上百条。ＧＡＧ是由二糖重复单位（氨基己糖、己糖醛...     

罢生植物根转的丛枝菌根真菌Ⅱ.

本文主要报道了野生植物根围Ｇｌｏｍｕｓ属的１７个种,聚球囊霉Ｇ．ａｇｇｒｅｇａｔｕｍ Ｓｃｈｅｎｃｈ＆Ｓｍｉｔｈ。苏格兰球囊霉Ｇ．ｃａｌｅｄｏｎｉｕｍ（Ｎｉｃｏｌ．＆Ｇｅｒｄ．）Ｔｒａｐｐｅ＆Ｇｅｒｄ,近明球囊霉Ｇ．ｃｌａｒｏｉｄｕｅｕｍＳｃｈｅｎｃｋ＆Ｓｍｉｔｈ,明球囊霉Ｇ．ＣＬＡＲＵＭ ｎＩＣＯＬＳＯＮ＆Ｓｃｈｅｎｃｋ,缩球囊霉Ｇ．ｃｏｎｓｔｒｉｃｔｕｍ Ｔｒａｐｐｅ,透光球囊霉Ｇ．ｆａｓｃ     

丝瓜的三聚体G蛋白的初步研究

根据拟南芥（ａｒａｂｉｄｏｐｓｉｓｔｈａｈｌｉａｎａ）ＧＰＡ１的保守区段Ａ设计一对特异引物（５′ｃｔｇｇｇｇａａｔｃｔｇｇａａａａｔｃ３′,５′ｃａｃａｇｃｔｇｔａｃａｃｃｔｃａａａｃ３′）通过ＰＣＲ从丝瓜核基因组中扩增植物的三聚体Ｇ蛋白α亚基编码基因,获得了２个片段（ＬＦＧ１,ＬＦＧ２）,并已克隆和测序（已在ＥＭＢＬ数据库中登记,登记号为：ｙ１５２７０,ｙ１５２７１）．序列分析表明ＬＦＧ１和ＬＦＧ２分别由１５１５ｂｐ和７３２ｂｐ构成,都含有三聚体Ｇ蛋白α亚基编码基因的保守区段Ａ,但也都含有内含子．根据片段的大小及ＰＣＲ的特性,ＬＦＧ１可能是丝瓜三聚体Ｇ蛋白α亚基编码基因上的片段．     

A New Journal From China:Genomics ,Proteomics & Bioinformatics

Ｇｅｎｏｍｉｃｓ ,Ｐｒｏｔｅｏｍｉｃｓ＆Ｂｉｏｉｎｆｏｒｍａｔｉｃｓ (ＧＰＢ)ｉｓｓｐｏｎｓｏｒｅｄｂｙｔｈｅＢｅｉｊｉｎｇＧｅｎｏｍｉｃｓＩｎｓｔｉｔｕｔｅ (ＢＧＩ) ,ｔｈｅＩｎｓｔｉ ｔｕｔｅｏｆＧｅｎｅｔｉｃｓａｎｄＤｅｖｅｌｏｐｍｅｎｔａｌＢｉｏｌｏｇｙｉｎＣｈｉｎｅｓｅＡｃａｄｅｍｙｏｆＳｃｉｅｎｃｅｓ (ＣＡＳ) ,ａｎｄｔｈｅＧｅｎｅｔｉｃｓＳｏｃｉｅｔｙｏｆＣｈｉｎａ ,ａｎｄｐｕｂｌｉｓｈｅｄｂｙｔｈｅＳｃｉｅｎｃｅＰｒｅｓｓ ,Ｂｅｉｊｉｎｇ .Ｔｈｅｆｉｒｓｔｉｓｓｕｅｏｆｔｈｉｓｑｕａ…     

Mechanism of aluminium-induced porphyrin synthesis in bacteria

In previous studies, aluminium was found to retard bacterial growth and enhance porphyrin formation in Arthrobacter aurescens RS-2. The aim of this study was to establish the mechanism of action of aluminium which leads to increased porphyrin production. Cultures of Arthrobacter aurescens RS-2 were incubated in the absence and presence of 0.74 mm aluminium. After 6 and 24 h of incubation, various parameters of the haem biosynthetic pathway were determined. After 6 h of incubation with aluminium, the activities of the enzymes aminolevulinate synthase (ALAS), aminolevulinate dehydratase (ALAD), porphobilinogen deaminase (PBGD) and uroporphyrinogen decarboxylase (UROD) were increased by 120, 170, 190 and 203%, respectively, while that of ferrochelatase (FC) was found to be unchanged. However, after 24 h of incubation, no change in the activities of ALAS and ALAD was noted, while an about 2-fold increase in PBGD and UROD activities were observed. FC activity was decreased by 63%. It was concluded that aluminium exerts its effect by inducing the enzymes PBGD and UROD rather than by a direct or indirect effect on ALAS. Its effect on the final step in the haem biosynthetic pathway is discussed.     

The influence of carbamazepine on the heme biosynthetic pathway

Carbamazepine, a drug which is widely used in neurological diseases, has a porphyrogenic effect in chick embryo liver cells in culture. It increased the concentration of cellular porphyrins by 80-fold and delta-aminolevulinate synthase activity by 4-fold. The increase in the accumulation of porphyrins preceded that of ALAS activity. Measurements of the activities of aminolevulinate dehydrase, porphobilinogen deaminase, and uroporphyrinogen decarboxylase showed that C inhibits UROD up to nearly 50% and PBGD activity up to 20%, but does not affect the activity of ALAD. The pattern of accumulation of porphyrins, mainly uro- and heptacarboxylporphyrin, is compatible with an inhibition of UROD. We may, therefore, conclude that the porphyrogenic effect of C in monolayers of chick embryo liver cells is the result of its inhibitory effect on the activity of UROD.     

Influence of cesium on tetrapyrrole biosynthesis in etiolated and greening barley leaves

Cesium chloride (CsCl) treatment of greening primary leaves of barley for 8 h inhibited chlorophyl] accumulation in a concentration-dependent manner and led to the accumulation of excessive amounts of uroporphyrin(ogen) III (URO[gen]) and to a minor extent of heptacarboxylporphyrin(ogen). When dark-grown leaves were incubated with CsCl, accumulation of URO(gen) was observed only after feeding of the tetrapyrrole precursor 5-aminolevulinic acid. Western blot analysis showed no apparent difference in content of uroporphyrinogen decarboxylase (EC 4.1.1.37, UROD) or selected proteins involved in tetrapyrrole biosynthesis in extracts of CsCl-incubated (15 m M ) versus control leaves. UROD activity was drastically decreased upon CsCl treatment in leaves incubated in the dark or in the light (44 and 86%, respectively). Selected preceding enzymes of the tetrapyrrole biosynthetic pathway, 5-aminolevulinic acid dehydratase (EC 4.2.1.24, ALAD) and porphobilinogen deaminase (EC 4.3.1.8, PBGD), were influenced only to a minor extent under standard incubation conditions (15 m M CsCl). Furthermore, the ALA synthesizing capacity did not differ in leaves incubated with and without Cs⁻ cations. UROD activity of crude homogenates from control plants and after partial purification was reduced to 56 and 80%, respectively, upon addition of 10 m M CsCl. Equal concentrations of KCl were not inhibitory. Enzyme assays of the same barley extract in the presence of CsCl yielded no effect on ALAD and a minor loss of PBGD activity. The initial visible cytotoxic effect of CsCl appeared to be a selective inhibition of UROD resulting in accumulation of photosensitizing URO (gen). Consequences of the diminished UROD activity on early steps of the tetrapyrrole biosynthesis and its functional and regulatory significance for the porphyrin synthesis are discussed.     

The common origins of the pigments of life—early steps of chlorophyll biosynthesis

The complex pathway of tetrapyrrole biosynthesis can be dissected into five sections: the pathways that produce 5-aminolevulinate (the C-4 and the C-5 pathways), the steps that transform ALA to uroporphyrinogen III, which are ubiquitous in the biosynthesis of all tetrapyrroles, and the three branches producing specialized end products. These end products include corrins and siroheme, chlorophylls and hemes and linear tetrapyrroles. These branches have been subjects of recent reviews. This review concentrates on the early steps leading up to uroporphyrinogen III formation which have been investigated intensively in recent years in animals, in plants, and in a wide range of bacteria.Abbreviations ALA 5-aminolevulinic acid - ALAS 5-aminolevulinic acid synthase - GR glutamyl-tRNA reductase - GSA glutamate-1-semialdehyde - GSAT glutamate-1-semialdehyde aminotransferase - HMB hydroxymethylbilane - PBG porphobilinogen - PBGD porphobilinogen deaminase - PBGS porphobilinogen synthase - URO uroporphyrin - URO'gen uroporphyrinogen - US uroporphyrinogen III synthase     

Production of uroporphyrinogen III, which is the common precursor of all tetrapyrrole cofactors, from 5-aminolevulinic acid by Escherichia coli expressing thermostable enzymes

Uroporphyrinogen III (urogen III) was produced from 5-aminolevulinic acid (ALA), which is a common precursor of all metabolic tetrapyrroles, using thermostable ALA dehydratase (ALAD), porphobilinogen deaminase (PBGD), and urogen III synthase (UROS) of Thermus thermophilus HB8. The UROS-coding gene (hemD ₂ ) of T. thermophilus HB8 was identified by examining the gene product for its ability to produce urogen III in a coupled reaction with ALAD and PBGD. The genes encoding ALAD, PBGD, and UROS were separately expressed in Escherichia coli BL21 (DE3). To inactivate indigenous mesophilic enzymes, the E. coli transformants were heated at 70 °C for 10 min. The bioconversion of ALA to urogen III was performed using a mixture of heat-treated E. coli transformants expressing ALAD, PBGD, and UROS at a cell ratio of 1:1:1. When the total cell concentration was 7.5 g/l, the mixture of heat-treated E. coli transformants could convert about 88 % 10 mM ALA to urogen III at 60 °C after 4 h. Since eight ALA molecules are required for the synthesis of one porphyrin molecule, approximately 1.1 mM (990 mg/l) urogen III was produced from 10 mM ALA. The present technology has great potential to supply urogen III for the biocatalytic production of vitamin B₁₂.     

Unique properties of Plasmodium falciparum porphobilinogen deaminase

The hybrid pathway for heme biosynthesis in the malarial parasite proposes the involvement of parasite genome-coded enzymes of the pathway localized in different compartments such as apicoplast, mitochondria, and cytosol. However, knowledge on the functionality and localization of many of these enzymes is not available. In this study, we demonstrate that porphobilinogen deaminase encoded by the Plasmodium falciparum genome (PfPBGD) has several unique biochemical properties. Studies carried out with PfPBGD partially purified from parasite membrane fraction, as well as recombinant PfPBGD lacking N-terminal 64 amino acids expressed and purified from Escherichia coli cells (DeltaPfPBGD), indicate that both the proteins are catalytically active. Surprisingly, PfPBGD catalyzes the conversion of porphobilinogen to uroporphyrinogen III (UROGEN III), indicating that it also possesses uroporphyrinogen III synthase (UROS) activity, catalyzing the next step. This obviates the necessity to have a separate gene for UROS that has not been so far annotated in the parasite genome. Interestingly, DeltaPfP-BGD gives rise to UROGEN III even after heat treatment, although UROS from other sources is known to be heat-sensitive. Based on the analysis of active site residues, a DeltaPfPBGDL116K mutant enzyme was created and the specific activity of this recombinant mutant enzyme is 5-fold higher than DeltaPfPBGD. More interestingly, DeltaPfPBGDL116K catalyzes the formation of uroporphyrinogen I (UROGEN I) in addition to UROGEN III, indicating that with increased PBGD activity the UROS activity of PBGD may perhaps become rate-limiting, thus leading to non-enzymatic cyclization of preuroporphyrinogen to UROGEN I. PfPBGD is localized to the apicoplast and is catalytically very inefficient compared with the host red cell enzyme.     

A knock-in mouse model of congenital erythropoietic porphyria

Congenital erythropoietic porphyria (CEP) is a recessive autosomal disorder characterized by a deficiency in uroporphyrinogen III synthase (UROS), the fourth enzyme of the heme biosynthetic pathway. The severity of the disease, the lack of specific treatment except for allogeneic bone marrow transplantation, and the knowledge of the molecular lesions are strong arguments for gene therapy. An animal model of CEP has been designed to evaluate the feasibility of retroviral gene transfer in hematopoietic stem cells. We have previously demonstrated that the knockout of the Uros gene is lethal in mice (Uros(del) model). This work describes the achievement of a knock-in model, which reproduces a mutation of the UROS gene responsible for a severe UROS deficiency in humans (P248Q missense mutant). Homozygous mice display erythrodontia, moderate photosensitivity, hepatosplenomegaly, and hemolytic anemia. Uroporphyrin (99% type I isomer) accumulates in urine. Total porphyrins are increased in erythrocytes and feces, while Uros enzymatic activity is below 1% of the normal level in the different tissues analyzed. These pathological findings closely mimic the CEP disease in humans and demonstrate that the Uros(mut248) mouse represents a suitable model of the human disease for pathophysiological, pharmaceutical, and therapeutic purposes.     

A molecular study of congenital erythropoietic porphyria in cattle

Previous studies have shown that congenital erythropoietic porphyria (CEP) in cattle is caused by an inherited deficiency of the enzyme uroporphyrinogen III synthase (UROS) encoded by the UROS gene. In this study, we have established the pedigree of an extended Holstein family in which the disease is segregating in a manner consistent with autosomal recessive inheritance. Biochemical analyses demonstrated accumulation of uroporphyrin, thus confirming that it is indeed insufficient activity of UROS which is the cause of the disease. We have therefore sequenced all nine exons of UROS in affected and non-affected individuals without detecting any potential causative mutations. However, a single nucleotide polymorphism (SNP) located within the spliceosome attachment region in intron 8 of UROS is shown to segregate with the disease allele. Our study supports the hypothesis that CEP in cattle is caused by a mutation affecting UROS; however, additional functional studies are needed to identify the causative mutation.     

Congenital erythropoietic porphyria with two mutations of the uroporphyrinogen III synthase gene (Cys73Arg, Thr228Met)

Congenital erythropoietic porphyria (CEP) is an autosomal recessive inborn error of metabolism that results from the markedly deficient activity of uroporphyrinogen III synthase (UROS). We describe a 14-year-old girl with red urine since infancy, progressive blistering and scarring of the skin, and moderate hemolytic anemia. After years of skin damage, her face is mutilated; she has a bald patch on the scalp, hypertrichosis of the neck, areas of skin darkening, and limited joint movements of the hands. Total urine excretion and fecal total porphyrin were both markedly raised above normal levels. Sequencing of the UROS gene identified two mutations causing CEP (Cys73Arg, Thr228Met). The patient lesions are progressing. Bone marrow transplantation and/or gene therapy are proposed as the next steps in her treatment. In brief, we describe a CEP with confirmed two pathogenic mutations, severe phenotype and discuss the various treatment options available.