Reciprocal regulation of Δ 1-pyrroline-5-carboxylate synthetase and proline dehydrogenase genes controls proline levels during and after osmotic stress in plants

Plants generally accumulate free proline under osmotic stress conditions. Upon removal of the osmotic stress, the proline levels return to normal. In order to understand the mechanisms involved in regulating the levels of proline, we cloned and characterized a proline dehydrogenase (PDH) cDNA from Arabidopsis thaliana (AtPDH). The 1745?bp cDNA contains a major open reading frame encoding a peptide of 499 amino acids. The deduced amino acid sequence has high homology with both Saccharomyces cerevisiae and Drosophila melanogaster proline oxidases and contains a putative mitochondrial targeting sequence. When expressed in yeast, the AtPDH cDNA complemented a yeast put1 mutation and exhibited proline oxidase activity. We also determined the free proline contents and the Δ¹-pyrroline-5-carboxylate synthetase (P5CS) and PDH mRNA levels under different osmotic stress and recovery conditions. The results demonstrated that the removal of free proline during the recovery from salinity or dehydration stress involves an induction of the PDH gene while the activity of P5CS declines. The reciprocal regulation of P5CS and PDH genes appears to be a key mechanism in the control of the levels of proline during and after osmotic stress. The PDH gene was also significantly induced by exogenously applied proline. The induction of PDH by proline, however, was inhibited by salt stress.     

壳多糖酶研究的概况及最新进展

壳多糖酶专一性水解壳多糖中的β－1,4糖苷键,在自然界的碳循环中具有极其重要的意义．壳多糖酶分布广泛,功能多样,在真菌生长发育、植物抗真菌感染等生理过程中壳多糖酶均发挥重要作用．文章概述了壳多糖酶研究的现状及最新进展,介绍了壳多糖酶的分布、理化性质、催化性质、在细胞中的定位及其调控;简介了近年来真菌和植物壳多糖酶研究的动态及最新进展.     

药物对病毒感染培养心肌细胞Ca~（2＋）内流及CVB_3－RNA复制的影响

应用放射性同位素￣（４５）Ｃａ￣（２＋）示踪技术及光敏生物素标记ｃＤＮＡ探针杂交方法,观察了维拉帕米、黄芪、地塞米松等药物对感染柯萨奇Ｂ＿３病毒（ＣＶＢ＿３）的大鼠培养心肌细胞Ｃａ￣（２＋）内流及细胞中ＣＶＢ＿３－ＲＮＡ复制的作用。结果发现：在病毒感染４８ｈ,上述三药均可显著减少感染细胞及正常对照的心肌Ｃａ￣（２＋）内流（Ｐ＜０．０１和Ｐ＜０．０５）;若在病毒感染后即加入上述药物,经４８ｈ培养后,黄芪组细胞中的ＣＶＢ＿３－ＲＮＡ含量显著少于病毒对照组（Ｐ＜０．００１）,维拉帕米则显著增加（Ｐ＜０．０１）．而地塞米松对其无影响。提示黄芪与地塞米松具有一种与维拉帕米相似的减少病毒感染心肌Ｃａ￣（２＋）内流的作用,有可能减轻感染细胞的继发性Ｃａ￣（２＋）损伤;但三种药物对感染细胞中病毒核酸的复制有不同作用,可作为临床治疗病毒性心肌炎时的参考。     

Adeno-associated virus type 2-mediated transfer of ecotropic retrovirus receptor cDNA allows ecotropic retroviral transduction of established and primary human cells.

The cellular receptors that mediate binding and internalization of retroviruses have recently been identified. The concentration and accessibility of these receptors are critical determinants in accomplishing successful gene transfer with retrovirus-based vectors. Murine retroviruses containing ecotropic glycoproteins do not infect human cells since human cells do not express the receptor that binds the ecotropic glycoproteins. To enable human cells to become permissive for ecotropic retrovirus-mediated gene transfer, we have developed a recombinant adeno-associated virus type 2 (AAV) vector containing ecotropic retroviral receptor (ecoR) cDNA under the control of the Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter (vRSVp-ecoR). Established human cell lines, such as HeLa and KB, known to be nonpermissive for murine ecotropic retroviruses, became permissive for infection by a retroviral vector containing a bacterial gene for resistance to neomycin (RV-Neo(r)), with a transduction efficiency of up to 47%, following transduction with vRSVp-ecoR, as determined by the development of colonies that were resistant to the drug G418, a neomycin analog. No G418-resistant colonies were present in cultures infected with either vRSVp-ecoR or RV-Neo(r) alone. Southern and Northern blot analyses revealed stable integration and long-term expression, respectively, of the transduced murine ecoR gene in clonal isolates of HeLa and KB cells. Similarly, ecotropic retrovirus-mediated Neo(r) transduction of primary human CD34+ hematopoietic progenitor cells from normal bone marrow was also documented, but only following infection with vRSVp-ecoR. The retroviral transduction efficiency was approximately 7% without prestimulation and approximately 14% with prestimulation of CD34+ cells with cytokines, as determined by hematopoietic clonogenic assays. No G418-resistant progenitor cell colonies were present in cultures infected with either vRSVp-ecoR or RV-Neo(r) alone. These results suggest that sequential transduction of primary human cells with two different viral vectors may overcome limitations encountered with a single vector. Thus, the combined use of AAV- and retrovirus-based vectors may have important clinical implications for ex vivo and in vivo human gene therapy.     

磷脂模型膜的时间分辨纳秒荧光光谱的研究

叙述了用时间分辨率纳秒荧光技术研究大豆磷脂或合成磷脂模型膜（脂质体）的物理状态变化与荧光寿命以及时间分辨各向异性测量参数变化相关性的实验结果。用荧光探剂ＭＣ５４０标记模型膜的结果表明,荧光寿命（τ）的变化与脂质／探剂比例、磷脂组成、磷脂的极性头部、脂肪酸酰链长度等有关。用ＤＰＨ（１,６－ｄｉｐｈｅｎｙｌ－１,３,５－ｈｅｘａｔｒｉｅｎｅ）标记磷脂模型膜的时间分辨各向异性分析结果表明,序参数（Ｓ）、     

Purification of chicken brain choline acetyltransferase

Chicken brain choline acetyltransferase was purified to homogeneity using ammonium sulfate fractionation, followed by chromatography on DEAE-Sephadex (A-25), hydroxyapatite, Sephadex G-150, immunoabsorption and Sepharose-CoA columns. A purification of 3500-fold was achieved and the final preparation had a specific activity of 2:32 μmol acetylcholine formed per minute per milligram protein. The purified chicken choline acetyltransferase migrated as a single band on polyacrylamide gel electrophoresis in the presence and absence of sodium deodecyl sulfate. The native enzyme, with a molecular weight of 67,000 daltons, consists of two subunits of identical molecular weight. Chicken choline acetyltransferase has a sharp pH optimum of 7.4. It is activated by sodium chloride and potassium chloride but inhibited by cupric ion and N-ethylmaleimide.     

两株粗糙型沙门氏菌对小鼠抗异源G~-杆菌攻击的免疫保护研究

本文对引自日本的一株粗糙化学型变异株明尼苏达沙门氏菌Re595(J)和引自美国的一株Re595(A)对小鼠异源性G~-杆菌主动和被动保护作用进行了比较。结果表明,两株菌对异源G~-杆菌的大肠杆菌和变形杆菌攻击均有良好的保护作用,但Re595(J)对抗肺炎克雷伯氏杆菌攻击的保护作用明显优于Re595(A),而Re595(A)抗绿脓杆菌攻击的保护作用则明显优于Ke595(J)。表明两株Re595的免疫原存在着差异。     

菌种与品种最佳组合的选育及增产效果

本文通过多年多点的田间接种试验,分析了大豆根瘤菌C_(33)(系VSDA_(110)的突变株)的增产效应.在合丰25号大豆接种C_(33)取得明显的增产效果,两年平均比CK增产28.55%,比61A76增产24.5%;C_(33)接种在其他品种以及在不同类型和肥力的土壤上增产效果也均高于CK和61A76,说明大豆根瘤菌C_(33)比当前生产上应用的61A76更具有广谱性和高效性.     

Fulvic acid disturbs processing of procollagen II in articular cartilage of embryonic chicken and may also cause Kashin-Beck disease.

Kashin-Beck disease is an endemic osteoarthropathy in China which may lead to skeletal deformation and dwarfism. We have analysed articular cartilage from two patients and found an accumulation of the precursor molecule, pro-pN-collagen II (pN, peptide attached at the amino-terminus) which was not present in extracts of control fetal cartilage. In addition, collagen II isolated from the same tissue by limited pepsin digestion had a decreased electrophoretic mobility, increased proline hydroxylation and decreased thermal stability. Previously, a genetic defect in pro-pN-collagen-I processing has been described in calf and sheep (dermatosparaxis) and man (Ehlers-Danlos, type VII) which caused an extreme fragility of the skin [Lenaers, A., Ansay, M., Nusgens, B.V. & Lapière, C.M. (1971) Eur. J. Biochem. 23, 533-541; Helle, O. & Nes, N.J. (1972) Acta Vet. Scand. 13, 443-445; Lichtenstein, J.R., Martin, G.R., Kohn, L.D., Byers, P.H. & McKusick, V.A. (1973) Science 182, 298-300]. Accordingly, one may assume that the impaired conversion of pro-pN-collagen II to collagen II and the structural alteration of collagen II, presumably caused by fulvic acid and other environmental factors, play an important role in the pathogenesis of Kashin-Beck disease.