杭州石荠苧生态学特性研究

 杭州石荠苧(Mosla hangchowensis)的种子完全靠风传播,但由于种子大,传播距离不远;种子在冬季休眠,春天(2月末3月初)萌发,种子萌发率很低,尤其是水选上层种子,主要原因是质量差。杭州石荠苧的营养期从3月初到8月上旬,株高在8月中旬以前基本为匀速增加,早期生长极为缓慢。形态和生殖力的环境可塑性极强,自然生长的植株冠幅变动在4～5616cm2之间。杭州石荠苧在自然生境中有时形成单优群落,通常与其它植物伴生。由于早期生长慢,限制了其在群落中的竞争能力,在土壤条件好的地方绝大部分被排挤掉,只是由于其极强的耐旱能力才在高温、干旱、土少的生境中得以存活。 将同属不濒危的华荠苧与之比较,其种子小于杭州石荠苧,但萌发率却高于杭州石荠苧。华荠苧的植株较矮,花色不如杭州石荠苧鲜艳,同在路边生长,不像杭州石荠苧那样容易被人采摘;华荠苧的根较杭州石荠苧的根深,抗雨水冲刷能力较强。华荠苧在自然生境中植株投入生殖的比例大于杭州石荠苧。     

Allosteric activation mechanism of the alpha1beta2gamma2 gamma-aminobutyric acid type A receptor revealed by mutation of the conserved M2 leucine

A conserved leucine residue in the midpoint of the second transmembrane domain (M2) of the ligand-activated ion channel family has been proposed to play an important role in receptor activation. In this study, we assessed the importance of this leucine in the activation of rat alpha1beta2gamma2 GABA receptors expressed in Xenopus laevis oocytes by site-directed mutagenesis and two-electrode voltage clamp. The hydrophobic conserved M2 leucines in alpha1(L263), beta2(L259), and gamma2(L274) subunits were mutated to the hydrophilic amino acid residue serine and coexpressed in all possible combinations with their wild-type and/or mutant counterparts. The mutation in any one subunit decreased the EC(50) and created spontaneous openings that were blocked by picrotoxin and, surprisingly, by the competitive antagonist bicuculline. The magnitudes of the shifts in GABA EC(50) and picrotoxin IC(50) as well as the degree of spontaneous openings were all correlated with the number of subunits carrying the leucine mutation. Simultaneous mutation of the GABA binding site (beta2Y157S; increased the EC(50)) and the conserved M2 leucine (beta2L259S; decreased the EC(50)) produced receptors with the predicted intermediate agonist sensitivity, indicating the two mutations affect binding and gating independently. The results are discussed in light of a proposed allosteric activation mechanism.     

Dual emission fluorescent silver nanoclusters for sensitive detection of the biological coenzyme NAD+/NADH

Fluorescent silver nanoclusters (Ag NCs) displaying dual-excitation and dual-emission properties have been developed for the specific detection of NAD⁺ (nicotinamide adenine dinucleotide, oxidized form). With the increase of NAD⁺ concentrations, the longer wavelength emission (with the peak at 550 nm) was gradually quenched due to the strong interactions between the NAD⁺ and Ag NCs, whereas the shorter wavelength emission (peaking at 395 nm) was linearly enhanced. More important, the dual-emission intensity ratio (I₃₉₅/I₅₅₀), fitting by a single-exponential decay function, can efficiently detect various NAD⁺ levels from 100 to 4000 μM, as well as label NAD⁺/NADH (reduced form of NAD) ratios in the range of 1–50.     

人GDNF基因在昆虫细胞中的高效表达

应用昆虫杆状病毒表达系统在昆虫细胞Ｔｎ－５Ｂ１－４中高效表达了人胶质细胞源性神经营养因子（ＧＤＮＦ）,ＰＡＧＥ分析表达量占细胞可溶性蛋白质的３０％左右,表达产物经亲和层析纯化后纯度达８０％以上,活性研究表明,昆虫细胞表达的ＧＤＮＦ蛋白能显著促进多巴胺能神经元的存活,此研究为进一步研究ＧＤＮＦ结构与功能打下了良好的基础。     

洪湖鱼类多样性研究

根据１９９２～１９９３年洪湖鱼类资源调查，结合１９５９年和１９８２年的历史资料，分析了洪湖鱼类物种多样性和群落结构多样性的变化．结果表明，洪湖鱼类从江湖阻隔初期的湖泊定居性鱼类为主、江湖洄游性鱼类和少数河海洄游性鱼类亦有一定的产量、大型和中型鱼类占较大比例，逐渐演变成以鲫占绝大多数的“众小”局面；鱼类群落的营养结构从草食性种类的比例高达２３％～３５％、肉食性种类比例波动于１０％～２０％之间，逐渐演变成以鲫为主的杂食性鱼类占绝大多数，草食性种类基本上从湖内消失；种类组成从阻隔前的推测应不下１００种演变成５０余种，且实际能在洪湖生息的只有３０余种；多样性指数、均匀度指数均有不同程度的降低．     

Molecular analysis of kanamycin and viomycin resistance in Mycobacterium smegmatis by use of the conjugation system.

We examined the molecular mechanisms of resistance to kanamycin and viomycin in Mycobacterium smegmatis. All of the M. smegmatis strains with high-level kanamycin resistance had a nucleotide substitution from A to G at position 1389 of the 16S rRNA gene (rrs). This position is equivalent to position 1408 of Escherichia coli, and mutation at this position is known to cause aminoglycoside resistance. Mutations from G to A or G to T at position 1473 of the M. smegmatis rrs gene were found in viomycin-resistant mutants which had been designated vicB mutants in our earlier studies. Using the M. smegmatis conjugation system, we confirmed that these mutations indeed contributed to kanamycin and viomycin resistance, and kanamycin susceptibility was dominant over resistance in a heterogenomic strain. Additional experiments showed that three of four Mycobacterium tuberculosis strains with high-level kanamycin resistance had a mutation from A to G at position 1400, which was equivalent to position 1389 of M. smegmatis.     

Effect of culture conditions on the whole cell activity of recombinant <Emphasis Type="Italic">Escherichia coli</Emphasis> expressing periplasmic organophosphorus hydrolase and cytosolic GroEL/ES chaperone

Specific whole cell activity strongly affects sensitivity and detection limit of whole cell-based biosensors. Previously, we developed recombinant Escherichia coli coexpressing periplasmic organophosphorus hydrolase (OPH) and cytosolic chaperone GroEL-GroES (GroEL/ES). In present work, we investigated the effect of culture conditions on whole cell OPH activity. Especially, the whole cell OPH activity was significantly affected by the concentration of tetracycline that is an inducer for chaperone GroEL/ES. When cultured at 20°C for 31 h in M9 medium containing 1 mM IPTG, 50 ng/mL tetracycline, and 500 µM CoCl₂, the recombinant E. coli exhibited a specific whole cell OPH activity (U/OD₆₀₀) of ~0.55, which is 2.6-fold higher than that of recombinant E. coli cultured as previously described conditions. In addition, recombinant cells showed adequate storage stability for 1 week with 100% of original response. Finally, the improved activity and adequate stability in the whole cell biocatalyst will contribute to sensitivity, detection time, and stability of a whole cell-based biosensor for the detection of toxic organophosphates.     

Reusable biosorbents in capsules from zoogloea ramigera cells for cadmium removal

A biosorbent was prepared by immobilizing and culturing Zoogloea ramigera cells in calcium alginate capsules to high density. The biosorbent (the cell and its exopolysaccharide "Zooglan") along with the [calcium] alginate is known to be responsible for cadmium removal. The dry weight of the biosorbent reached 107 g/L after 3 days of cultivation and 220 g/L after 5 days based on the core volume of a 2.0-mm diameter capsule used. The biosorbents were completely contained in the core of the capsule where the cells grew preferentially near the shell of the capsules while the polymer distributed homogeneously in the core. The specific cadmium uptake by the capsule biosorbent was 1.9 mg/g adsorbent at an initial cadmium concentration of 3 mg/L. This is 1.24 times more than the specific cadmium uptake by the 1.8-mm beads prepared under a comparable condition. The capsules crosslinked with 1% triethylene tetramine and 1% glutamic dialdehyde solutions were superior to the uncrosslinked capsules in mechanical strength. The crosslinked capsules maintained their mechanical strength and adsorption/desorption capacity even after 30 cycles of repeated use. Copyright 1999 John Wiley & Sons, Inc.