银杏科研的主要成就和展望

前言银杏又名白果,是现存种子植物中最古老的孑遗植物,也是源于我国的特有珍贵树种。学者们一向把它视为植物界的奇珍,在植物分类学、树木学、果树栽培学和古植物学上占有特殊的位置,至今国内外植物学家、地质学家,以及果树栽培学家们,仍对它进行着深入细致的研究。银杏的科学奥秘将逐步被揭开。     

子宫颈糜烂病毒病因的探讨

491份宫颈拭子病毒分离结果表明:糜烂宫颈单纯疱疹病毒(HSV)分离阳性率(30.8％)是正常宫颈(2.6％)的11.8倍,用人干扰素治疗一个疗程后,病毒分离率下降至疗前的1/4.36例糜烂宫颈活体组织DNA分子杂交表明,乳头瘤病毒16型(HPV-16)阳性者占52.8和,HPV-18占17.9％,HPV-6B占28.1％,HPV-11占7.7％,251例宫颈糜烂患者经人(?)D型基因工程干扰素双盲对比治疗后,总有效率达93.8％,显效率达60％,分析临床疗效与HSV分离率的变化表明,临床有效病例中有35％(49/140)在治疗后病毒阴转,有57％在疗前疗后均未分离出HSV,有5％在疗前疗后保持阳性不变,有2.9％疗前阴性,疗后阳性,上述结果表明,HSV和HPV与慢性宫颈炎有一定关系。     

沙雷氏杆菌(Serratia marcescens)红素缩合酶的提取与稳定性的研究

 <正> 红素(Prodigiosin)是某些沙雷氏杆菌(Serratia marcescens)菌株所合成的一种三吡咯衍生物的红色色素,它对某些细菌,真菌和原虫具有抗生作用,因而引起人们的注意。在活菌体内,它由两个中间前体:2-甲基-3-戊基吡咯(MAP)和4-甲氧基-2,2’-双吡咯-5-甲醛(MBC)缩合而成。催化这一反应的酶,我们称之为红素缩合酶。除MAP外,某些人     

大鼠非致瘤四倍体细胞系的建立及细胞遗传学和酶学特征

从大鼠自然诱发的肉瘤细胞中,我们建立了一个四倍体细胞系(4n=84),命名为RC(ratcell)。它具有典型的成纤维细胞外形,能在玻璃表面贴壁生长,但不能生长在琼脂半固体培养基中。该细胞在含15％小牛血清的RPMI 1640培养基中生长良好,至今已连续繁殖112世代,细胞群体倍增时间约为15小时。染色体G-带分析表明,RC为整四倍体细胞,它的1条X染色体在第32至34区为均染区。RC细胞核仁组织者(NORs)活性显然比大鼠二倍体细胞NORs活性的加倍还高(P<0.001)。这个具有非常高NORs活性的RC细胞系对于研究细胞18S+28S rRNA基因转录活性的调控、基因表达与基因剂量关系有一定的意义。RC细胞还有异常高的磷酸酯酶活性,而且它的同工酶谱也与大鼠肌肉细胞明显不同。体内接种实验和扫描电镜的观察表明,RC是非致瘤细胞。RC细胞各号染色体的C-带图样与大鼠二倍体细胞无明显的差异。     

Electrophoretic behavior of the H+-ATPase proteolipid from bovine heart mitochondria

The proteolipid subunit of H⁺-ATPase was labeled by [¹⁴C]N,N-dicyclohexylcarbodiimide in bovine heart mitochondria. The radioactive labeling was followed using various systems of sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). When using discontinuous SDS-PAGE (Laemmli, U.K., 1970,Nature (London)227, 680–685) a monomeric (Mr 7600±1500) and a dimeric form (Mr 17,800±1200) of the proteolipid were detected, while only the monomeric form was found on urea (8 M) containing gels (SDS-PAGE according to Laemmli; or Swank, R. T., and Munkers, K. D., 1971,Anal. Biochem. 39, 462–477). When using SDS-PAGE with Na-P_i buffer (Weber, K., and Osborn, M., 1969,J. Biol. Chem. 244, 4406–4442), only a dimeric form of the proteolipid (Mr 15,000±1000) was detected. Experimental data indicate that the different patterns of proteolipid separation are related to the presence of the two distinct proteolipid conformations in the SDS solution.     

重楼属植物的免疫血清学研究

以海南重楼(Paris dunniana),花叶重楼(P.marmorata),多叶重楼(P.polyphylla),凌云重楼(P.cronquistii),五指莲(P.axialis),滇重楼(Paris polyphylla var.yunnanensis)等六种重楼属植物的根茎为材料,提取分离其球蛋白组份作为抗原,免疫家兔得到相应的抗血清。通过免疫双扩散,聚丙烯酰胺凝胶电泳、免疫电泳、免疫吸收试验以及酶标免疫等方法,研究了重楼属上述六个种和变种的血清学行为以及他们之间的相互关系。在此基础上,用不加权算术平均对群法(UPGMA)经成聚运算,得到基于平均吸收相似系数(Sabs.mean.)和Jaccard's结合系数的两种表相图,这两种表相图的结果是一致的。花叶重楼与多叶重楼有较大的血清反应相似性,五指莲与滇重楼有最大的血清反应相似性。多型种P.polyphylla在形态上的较大变异与血清反应分析的结果是一致的,而且血清反应相似性分析支持了从形态分析得出的假设:Paris dunniana在重楼属的系统发生中是一比较原始的种。     

~(60)CO-γ射线诱发的德国蜚蠊精母细胞中心体畸变

德国蜚蠊经不同剂量的~(60)Co-γ射线照射,以光镜和电镜观察精母细胞巨大中心体结构和数目的变化,发现:γ射线可诱发各种类型中心体畸变,如棒状、断裂、不分离、超数和微小中心体等;畸变率随剂量增加而升高;畸变类型和辐射剂量有密切关系,低剂量(500—1000r)诱发较多的棒状中心体和中心体断裂、中等剂量(2000r)诱发较多的超数中心体和中心体不分离、高剂量(5000r)诱发较多的微小中心体,看来低剂量γ射线主要影响中心体结构,高剂量则对中心体的生长发育有明显的阻滞作用。本文还讨论了中心体畸变的可能产生机制及在遗传毒理研究中应用的前景。     

Microbial oxidation of methanol: Purification and properties of formaldehyde dehydrogenase from a Pichia sp. NRRL-Y-11328

An NAD⁺-linked, reduced glutathione-dependent formaldehyde dehydrogenase was purified to homogeneity from soluble extracts of methanol-grown yeast, Pichia sp. Formaldehyde and methylglyoxal are oxidized in the presence of NAD⁺ as an electron acceptor. NADP⁺ could not replace NAD⁺. Other straight chain aldehydes (C₂–C₆ tested), branched-chain aldehydes (e.g., isobutyaldehyde), aromatic aldehydes (e.g., salicylal-dehyde, benzaldehyde), glutyraldehyde, glyceraldehyde, glycoaldehyde, and glyoxal-dehyde tested were not oxidized by the purified formaldehyde dehydrogenase. The product of formaldehyde oxidation by purified enzyme was demonstrated to be S-for-mylglutathione by measuring the absorption at 240 nm due to the formation of thioester of formaldehyde and reduced glutathione. The K_m values for NAD⁺, formaldehyde, and reduced glutathione were 0.12, 0.31, and 0.16 mm, respectively, for the forward reaction at pH 8.0. The purified formaldehyde dehydrogenase also catalyzed the reduction of S-formylglutathione in the presence of NADH. Formate was not reduced by the purified enzyme. The K_m values for S-formylglutathione and NADH were 0.60 and 0.25 mm, respectively, for the reverse reaction at pH 6.0. Formaldehyde dehydrogenase has a molecular weight of 84,000 as determined by gel filtration and subunit molecular weight of 41,000 as determined by sodium dodecyl sulfate-gel electrophoresis. S-Formylglutathione, a product of formaldehyde oxidation, was oxidized by the partially purified formate dehydrogenase from Pichia sp. Formate dehydrogenase has a higher affinity toward S-formylglutathione (K_m value 1.8 mm) than toward formate (K_m value 25 mm). Antiserum prepared against the purified formaldehyde dehydrogenase from Pichia sp. NRRL-Y-11328 forms strong precipitin bands with isofunctional enzymes from methanol-grown Pichia pastoris NRRL-Y-7556 and Torulopsis candida Y-11419 and weak precipitin bands with Hansenula polymorpha NRRL-Y-2214. No cross-reaction was observed with isofunctional enzyme derived from methanol-grown Kloeckera sp.     

Epoxidation of Short-Chain Alkenes by Resting-Cell Suspensions of Propane-Grown Bacteria

Sixteen new cultures of propane-utilizing bacteria were isolated from lake water from Warinanco Park, Linden, N.J. and from lake and soil samples from Bayway Refinery, Linden, N.J. In addition, 19 known cultures obtained from culture collections were also found to be able to grow on propane as the sole carbon and energy source. In addition to their ability to oxidize n-alkanes, resting-cell suspensions of both new cultures and known cultures grown on propane oxidize short-chain alkenes to their corresponding 1,2-epoxides. Among the substrate alkenes, propylene was oxidized at the highest rate. In contrast to the case with methylotrophic bacteria, the product epoxides are further metabolized. Propane and other gaseous n-alkanes inhibit the epoxidation of propylene. The optimum conditions for in vivo epoxidation are described. Results from inhibition studies indicate that a propane monooxygenase system catalyzes both the epoxidation and hydroxylation reactions. Experiments with cell-free extracts show that both hydroxylation and epoxidation activities are located in the soluble fraction obtained after 80,000 × g centrifugation.     

Production of Methyl Ketones from Secondary Alcohols by Cell Suspensions of C2 to C4n-Alkane-Grown Bacteria

Nineteen new C₂ to C₄n-alkane-grown cultures were isolated from lake water from Warinanco Park, Linden, N.J., and from lake and soil samples from Bayway Refinery, Linden, N.J. Fifteen known liquid alkane-utilizing cultures were also found to be able to grow on C₂ to C₄n-alkanes. Cell suspensions of these C₂ to C₄n-alkane-grown bacteria oxidized 2-alcohols (2-propanol, 2-butanol, 2-pentanol, and 2-hexanol) to their corresponding methyl ketones. The product methyl ketones accumulated extracellularly. Cells grown on 1-propanol or 2-propanol oxidized both primary and secondary alcohols. In addition, the activity for production of methyl ketones from secondary alcohols was found in cells grown on either alkanes, alcohols, or alkylamines, indicating that the enzyme(s) responsible for this reaction is constitutive. The optimum conditions for in vivo methyl ketone formation from secondary alcohols were compared among selected strains: Brevibacterium sp. strain CRL56, Nocardia paraffinica ATCC 21198, and Pseudomonas fluorescens NRRL B-1244. The rates for the oxidation of secondary alcohols were linear for the first 3 h of incubation. Among secondary alcohols, 2-propanol and 2-butanol were oxidized at the highest rate. A pH around 8.0 to 9.0 was found to be the optimum for acetone or 2-butanone formation from 2-alcohols. The temperature optimum for the production of acetone or 2-butanone from 2-propanol or 2-butanol was rather high at 60°C, indicating that the enzyme involved in the reaction is relatively thermally stable. Metal-chelating agents inhibit the production of methyl ketones, suggesting the involvement of a metal(s) in the oxidation of secondary alcohols. Secondary alcohol dehydrogenase activity was found in the cell-free soluble fraction; this activity requires a cofactor, specifically NAD. Propane monooxygenase activity was also found in the cell-free soluble fraction. It is a nonspecific enzyme catalyzing both terminal and subterminal oxidation of n-alkanes.