Polyhydroxyalkanoate accumulation in Streptomyces coelicolor affected by SCO7613 gene region

Polyhydroxyalkanoate (PHA) is stored as an important carbon and energy source in bacterial cells. For biomedical applications, gram-positive bacteria can be better sources of PHAs, since they lack outer membrane lipopolysaccharide. Although gram-positive Streptomyces coelicolor A3(2) has been indicated as a high potential PHA producer, pha C gene that encodes the key enzyme PHA synthase in the metabolic pathway is not determined in its genome. BLAST search results of the GenBank database argued that SCO7613 could specify a putative polyhydroxyalkanoate synthase (PhaC) responsible for PHA biosynthesis. Deduced amino acid sequence of SCO7613 showed the presence of conserved lipase box like sequence, ⁵⁵⁵GASAG⁵⁵⁹, in which serine residue was present as the active nucleophile. Present study describes deletion of putative S. coelicolor pha C gene via PCR dependent method. We showed that SCO7613 is not an essential gene in S. coelicolor and its deletion affected PHA accumulation negatively although it is not ceased. Transcomplementation abolished the mutant phenotype, demonstrating that the decrease in PHA resulted from the deletion of SCO7613.     

组织酸化参与外周痛觉传递的离子通道机制

组织酸化可以导致痛觉的产生.初级感觉神经元可以通过离子通道来感受外周的组织酸化.已鉴定了几个离子通道家族可能参与了外周组织酸化的感受：a.酸敏感离子通道(ASICs)是可以被酸直接门控的阳离子通道;b.辣椒素受体(VR1)可被酸敏化,同时可被pH＜6.0直接激活;c.P2X2和P2X2/3受体通道反应被酸上调;d.TwIK相关的酸感受钾通道（TASK）是被酸关闭的双孔内向整流钾通道.这些通道被酸所调控的共同结果就是提高了神经元的兴奋性.因此,它们在介导了组织酸化所诱导的痛觉感受和传递中具有重要作用.     

环境因子对豆科共生固氮影响的研究进展

环境因子的限制一直是豆科植物一根瘤菌共生固氮体系没有在农业生产中充分发挥作用的重要原因之一。目前,研究涉及的环境因子主要行水分、矿质营养元素、温度、重金属、钠盐、CO2、土壤类型以及pH等。水分胁迫会导致豆科植物根瘤减少和固氮效率低下;矿质元素方面,除氮磷钾外,微量死素对固氮影响也很明显;不适的温度会对豆科植物的结瘤固氮产生一定的限制;重金属能从不同方面直接和间接地影响共生同氮,寻找适合作尾矿先锋植物的豆科植物是当前的一个研究热点。本文除详细阐述了这方面开展的研究以外,还浅析了这方而研究目前国内外存在的一些主要问题和发展趋势。     

一种新的从新疆穴居狼蛛中鉴定的细胞裂解肽

蜘蛛粗毒中富含生物活性物质,尤其以多肽类成分为主. 分离鉴定了新疆穴居狼蛛粗毒中一种新的细胞裂解肽,命名为LSTX-A1,其分子量为7 335.33,含有65个氨基酸残基,且羧基端残基酰胺化.研究了LSTX-A1的细胞裂解作用,结果显示,在100 μmol/L浓度引起42% 红细胞溶血.同时LSTX-A1具有抗肿瘤作用,能抑制HeLa细胞的增殖,其IC50剂量为22 μmol/L.     

Requirement of extracellular signal-regulated kinase/mitogen-activated protein kinase for long-term potentiation in adult mouse anterior cingulate cortex

Long-term potentiation (LTP) in the anterior cingulate cortex (ACC) is believed to be critical for higher brain functions including emotion, learning, memory and chronic pain. N-methyl-D-aspartate (NMDA) receptor-dependent LTP is well studied and is thought to be important for learning and memory in mammalian brains. As the downstream target of NMDA receptors, the extracellular signal-regulated kinase (ERK) in the mitogen-activated protein kinase (MAPK) cascade has been extensively studied for its involvement in synaptic plasticity, learning and memory in hippocampus. By contrast, the role of ERK in cingulate LTP has not been investigated. In this study, we examined whether LTP in ACC requires the activation of ERK. We found that P42/P44 MAPK inhibitors, PD98059 and U0126, suppressed the induction of cingulate LTP that was induced by presynaptic stimulation with postsynaptic depolarization (the pairing protocol). We also showed that cingulate LTP induced by two other different protocols was also blocked by PD98059. Moreover, we found that these two inhibitors had no effect on the maintenance of cingulate LTP. Inhibitors of c-Jun N-terminal kinase (JNK) and p38, other members of MAPK family, SP600125 and SB203850, suppressed the induction of cingulate LTP generated by the pairing protocol. Thus, our study suggests that the MAPK signaling pathway is involved in the induction of cingulate LTP and plays a critical role in physiological conditions.     

Generation of potent mouse monoclonal antibodies to self-proteins using T-cell epitope “tags”

Immunization of mice or rats with a "non-self" protein is a commonly used method to obtain monoclonal antibodies, and relies on the immune system''s ability to recognize the immunogen as foreign. Immunization of an antigen with 100% identity to the endogenous protein, however, will not elicit a robust immune response. To develop antibodies to mouse proteins, we focused on the potential for breaking such immune tolerance by genetically fusing two independent T-cell epitope-containing sequences (from tetanus toxin (TT) and diphtheria toxin fragment A (DTA)) to a mouse protein, mouse ST2 (mST2). Wild-type CD1 mice were immunized with three mST2 tagged proteins (Fc, TT and DTA) and the specific serum response was determined. Only in mice immunized with the T-cell epitope-containing antigens were specific mST2 serum responses detected; hybridomas generated from these mice secreted highly sequence-diverse IgGs that were capable of binding mST2 and inhibiting the interaction of mST2 with its ligand, mouse interleukin (IL)-33 (mIL-33). Of the hundreds of antibodies profiled, we identified five potent antibodies that were able to inhibit IL-33 induced IL-6 release in a mast cell assay; notably one such antibody was sufficiently potent to suppress IL-5 release and eosinophilia infiltration in an Alternaria alternata challenge mouse model of asthma. This study demonstrated, for the first time, that T-cell epitope-containing tags have the ability to break tolerance in wild-type mice to 100% conserved proteins, and it provides a compelling argument for the broader use of this approach to generate antibodies against any mouse protein or conserved ortholog.     

水分胁迫和氮肥对花生根系形态发育及叶片生理活性的影响

以"花育22号"花生为试验材料,在中度干旱胁迫和充足灌水两个水分条件下,分别设置不施氮肥(N0)、中氮(N1,90 kg·hm-2)、高氮(N2,180 kg·hm-2)3个施氮水平,研究不同土壤水分和氮肥条件对花生叶片生理活性及根系形态发育特征的影响.结果表明:与不施氮肥处理相比,两个水分条件下中氮处理均显著增加花生产量,但对收获指数无显著影响.干旱胁迫条件下,中氮处理对总根系生物量和总根长无显著影响,但显著增加花生总根系表面积;中氮和高氮处理均显著增加20~40 cm土层内根长和根系表面积,且高氮处理显著增加40 cm以下土层内根系生物量和根系表面积;施用氮肥显著提高叶片过氧化氢酶(CAT)和过氧化物酶(POD)活性,而丙二醛(MDA)含量随施氮量的增加而降低.正常供水条件下,施用氮肥显著降低了花生根系表面积和40 cm以下土层内根系生物量、根长和根系表面积,中氮处理可提高叶片保护酶活性.相关性分析表明,20~40 cm土层内根长和叶片超氧化物歧化酶(SOD)、CAT、POD活性与产量呈显著相关.