草鱼呼肠孤病毒RNA聚合酶基因的表达与产物纯化

草鱼呼肠孤病毒是引起草鱼出血病的主要病原,隶属于呼肠孤病毒科水生呼肠孤病毒属.序列分析表明,GCRV S2 片段长为3 877核苷酸,编码一个分子量为138kDa 的蛋白VP2,具有RNA聚合酶性质.为进一步了解该病毒 RNA聚合酶特性,本研究在对GCRV RNA聚合酶基因(GCRV-RdRp)保守区(约1.5kb)重组质粒pR/RRp高效表达的基础上,分别构建了编码GCRV RNA聚合酶保守区N端与C端部分基因的 pR/RRpN及pR/RRpC重组表达载体,并在原核细胞中获得成功表达.筛选的重组表达菌株经IPTG诱导培养,得到分子量分别为98kDa、103kDa的目的表达融合蛋白.Western blot分析表明,该表达产物与兔抗GCRV-VP2血清呈阳性反应.通过ProBond柱亲和层析,纯化了融合有6个组氨酸的重组表达产物,并获得约90%纯的目的蛋白.上述结果为GCRV RNA聚合酶特性分析提供了依据.     

Angiostatin production in cultivation of recombinant Pichia pastoris fed with mixed carbon sources

A recombinant strain of Pichia pastoris with a phenotype of Mut^S was used to produce angiostatin. Due to the low methanol consumption rate of this strain, both methanol and glycerol feedings, that produced oscillation in dissolved O₂ concentration, were used during the expression phase to improve cell growth and angiostatin expression. However, enhanced cell growth led to nitrogen limitation that suppressed further production of angiostatin, but addition of ammonia allowed angiostatin concentration to reach 108 mg l^–1 after an expression period of 96 h. The ratio of consumed glycerol to methanol of 1.5:1 (w/w) in the expression phase suggested that methanol played an important role in the metabolism of carbon sources.     

Wistar大鼠心脏自发性病变的病理学观察

目的了解Wistar大鼠心脏自发性病变发病情况,为长期致癌性研究、老年病学研究及毒性病理学提供背景资料。方法采用160只清洁级Wistar大鼠,雌雄各半,常规饲养,分别在9月龄、12月龄、18月龄、24月龄时处死40只大鼠,HE及Masson三色法染色,观察心脏的病理改变。结果 9月龄Wistar大鼠心脏未见明显病理改变;12月龄Wistar大鼠月龄心脏病变的发病率为2.5%（1/40）,表现为少数心肌细胞变性坏死伴少量以单核细胞为主的炎细胞浸润;18月龄大鼠心脏病变的发病率为57.5%（23/40）,表现为轻至中度心肌病,雄性发病率高于雌性。24月龄大鼠100%（40/40）出现不同程度的心肌病,并有2.5%（1/40）发生心内膜下纤维组织增生。Masson染色显示9月龄大鼠心脏血管周围及心脏瓣膜环下有少量胶原纤维,随年龄增长,血管周围及心脏瓣膜环下胶原纤维逐渐增多,并延伸入心肌细胞间。结论随年龄增长,大鼠心脏自发病变比率升高,主要病变为心肌病,偶尔可发生心内膜下纤维组织增生;胶原纤维沉积首先发生于血管周围及心脏瓣膜环下,随年龄增长而增多,可能与大鼠心肌病的的发生密切相关。     

Field trapping of male Phyllonorycter ringoniella using variable ratios of pheromone components

The sex pheromone of Phyllonorycter ringoniella (Matsumura) (Lepidoptera: Gracillariidae) has been identified to be a blend of (Z)‐10‐tetradecenyl acetate (Z10‐14:OAc) and E4,Z10‐tetradecadienyl acetate (E4,Z10‐14:OAc) in Japan, Korea, and China. However, the commercial product based on previous results is not attractive enough to be used for monitoring and controlling apple leafminer populations in the field. We re‐investigated the attractiveness of the two pheromone components, singly and in blends, in apple orchards in Shangdong and Shaanxi, the main apple‐growing provinces in China. Our results revealed that Z10‐14:OAc alone was not attractive to P. ringoniella male moths in the field, but E4,Z10‐14:OAc alone not only was strongly attractive but caught more males than any of the blends of Z10‐14:OAc and E4,Z10‐14:OAc tested. The most attractive blend ratios differed slightly for the two locations. No clear dose–response relationship was obtained for the 2:8 blend of Z10‐14:OAc and E4,Z10‐14:OAc. However, the dose–response field study of E4,Z10‐14:OAc alone showed that 1 mg per lure achieved the highest moth catch. These findings differ from the previous report of the best pheromone blend in China. Our data showed that E4,Z10‐14:OAc is the major component of the pheromone of P. ringoniella.     

Proton-dependent electron transfer from CuA to heme a and altered EPR spectra in mutants close to heme a of cytochrome oxidase

Eukaryotic cytochrome c oxidase (CcO) and homologous prokaryotic forms of Rhodobacter and Paraccocus differ in the EPR spectrum of heme a. It was noted that a histidine ligand of heme a (H102) is hydrogen bonded to serine in Rhodobacter (S44) and Paraccocus CcOs, in contrast to glycine in the bovine enzyme. Mutation of S44 to glycine shifts the heme a EPR signal from g(z) = 2.82 to 2.86, closer to bovine heme a at 3.03, without modifying other properties. Mutation to aspartate, however, results in an oppositely shifted and split heme a EPR signal of g(z) = 2.72/2.78, accompanied by lower activity and drastically inhibited intrinsic electron transfer from CuA to heme a. This intrinsic rate is biphasic; the proportion that is slow is pH dependent, as is the relative intensity of the two EPR signal components. At pH 8, the heme a EPR signal at 2.72 is most intense, and the electron transfer rate (CuA to heme a) is 10-130 s(-1), compared to wild-type at 90,000 s(-1). At pH 5.5, the signal at 2.78 is intensified, and a biphasic rate is observed, 50% fast (approximately wild type) and 50% slow (90 s(-1)). The data support the prediction that the hydrogen-bonding partner of the histidine ligand of heme a is one determinant of the EPR spectral difference between bovine and bacterial CcO. We further demonstrate that the heme a redox potential can be dramatically altered by a nearby carboxyl, whose protonation leads to a proton-coupled electron transfer process.     

混合暴露条件下近江牡蛎对重金属的积累与释放特征

选择近江牡蛎作为试验生物,研究了混合暴露条件下8种重金属在近江牡蛎体内的积累和释放特征.结果表明: 近江牡蛎对重金属Pb、Cu、Ni、Cd、Cr和Hg有很强的累积能力,可较好地指示溶液中的重金属浓度水平,但对重金属Zn和As的积累能力很小,不能真实反映溶液中重金属Zn和As含量的变化水平.在随后35 d的释放阶段,8种重金属在近江牡蛎体内的含量没有明显变化,表明近江牡蛎对重金属的释放能力较差.双箱动力学模型可较好地反映混合暴露条件下近江牡蛎对重金属的积累特征,但不适合对其释放特征进行描述.     

Two-dimensional crystallization of avidin on biotinylated lipid monolayers.

Two-dimensional crystals of avidin were obtained on mixed lipid monolayers containing biotinylated lipids (N-biotinyl-dipalmitoyl-L-alpha-phosphatidyl ethanolamine and dioleoyl phosphatidyl choline) by specific interaction. Image analysis of electron micrographs of these crystals revealed p2 symmetry with the unit cell parameters a = 66 +/- 2 A, b = 68 +/- 1 A, and gamma = 121 +/- 4 degrees. The projection map showed, at a resolution of about 27 A, that the four subunits within one avidin molecule are separated into two parts. Comparison between avidin and streptavidin reveals that avidin molecule binds to the lipid monolayer in an orientation similar to that of streptavidin.     

Identification of candidate chemosensory receptors in the antennal transcriptome of Tropidothorax elegans

Molecular Biology Reports - Chemosensory receptors in the dendritic membrane of olfactory cells are critical for the molecular recognition and discrimination of odorants. Tropidothorax elegans is a...