应用非光化学淬灭初始变化率作为浮游植物光保护能力指标的可行性

在非光化学淬灭(Non-Photochemical Quenching,NPQ)荧光诱导的早期(1 min),NPQ随时间的变化率(NPQ/t)可作为衡量浮游植物光保护能力的指标,与常用的传统指标NPQm(最大NPQ)相比,这一新指标具有更易标准化、测量效率更高且包含的信息更丰富的优点。通过实验测量和理论推导证明了NPQ/t的常数性质,并通过可控光照条件下的室内培养实验证明了应用NPQ/t反映浮游植物光保护能力的合理性和有效性:NPQ/t和NPQm、光保护色素浓度呈线性相关,高光强下培养的藻类其NPQ/t高于低光强下培养的藻类,不同藻种之间的NPQ/t:扁藻(Tetraselmis chui)牟氏角毛藻(Chaetoceros mulleri)叉鞭金藻(Dicrateria inornata)。这一方法解决了野外实测研究中传统指标难以满足结果标准化和快速测量等要求的问题,对于推动浮游植物光保护生态学研究具有重要意义。     

Serum resistance in Haemophilus parasuis SC096 strain requires outer membrane protein P2 expression

Haemophilus parasuis outer membrane protein P2 (OmpP2), the most abundant protein in the outer membrane, has been identified as an antigenic protein and a potential virulence factor. To study the precise function of OmpP2, an ompP2-deficient mutant (ΔompP2) of a H.?parasuis serovar 4 clinical strain SC096 was constructed by a modified natural transformation system. Compared with the wild-type SC096 strain, the ΔompP2 mutant showed a pronounced growth defect and exhibited significantly greater sensitivity to the bactericidal action of porcine and rabbit sera, whereas the complemented strain could restore the growth and serum resistance phenotypes. The results indicated that H.?parasuis OmpP2 from SC096 strain is an important surface protein involved in serum resistance.     

Correlations between the ages of Alnus host species and the genetic diversity of associated endosymbiotic Frankia strains from nodules

Nodule samples were collected from four alder species: Alnus nepalensis, A. si-birica, A. tinctoria and A. mandshurica growing in different environments on Gaoligong Mountains, Yunnan Province of Southwest China and on Changbai Mountains, Jilin Province of Northeast China. PCR-RFLP analysis of the IGS between nifD and nifK genes was directly applied to uncultured Frankia strains in the nodules. A total of 21 restriction patterns were obtained. The Frankia population in the nodules of A. nepalensis had the highest genetic diversity among all four Frankia populations; by contrast, the population in the nodules of A. mandshurica had the lowest degree of divergence; the ones in the nodules of A. sibirica and A. tinctoria were intermediate. A dendrogram, which was constructed based on the genetic distance between the restriction patterns, indicated that Frankia strains from A. sibirica and A. tinctoria had a close genetic relationship. Frankia strains from A. nepalensis might be the ancestor of Frankia strain     

Impacts of methamidophos,copper, and their combinations on bacterial community structure and function in black soil

The potential ecotoxicologial risks of methamidophos, copper, and their combinations on microbial community of black soil ecosystem in the Northeast China were assessed in species richness and structures by using 16S rDNA-PCR-DGGE analysis approach, and functional characteristics at community levels by using BIOLOG^GN system analysis method as well as two conventional methods(DHA and SIR). All results of DGGE banding fingerprint patterns(amplified by bacterial specific 16S rDNAV₃ high variable region universal primer) indicated that the species richness of bacterial community in tested soil was significantly decreased to different extents by using different concentrations of single methamidophos, copper, especially some of their combinations had worse effects than their corresponding single factors. In addition, the structures of soil bacterial community had been disturbed under all stresses applied in this study because of the enrichment of some species and the disappearance of other species from the bacterial community. The effects of the single factors with lower concentrations on the community structure were weaker than those with higher concentrations. Moreover, the bacterial community structures under the combined stresses of methamidophos and copper were significantly different from those of control and their corresponding single factors. The change of DHA and carbon source substrate utilizing fingerprint patterns based on BIOLOG^GNsystem were two relatively sensitive directors corresponding to the stress presented in this study. Between methamodophos and copper, there happened the significant joint-toxic actions when they were used in combination on DHA and carbon source substrate utilizing fingerprint patterns of soil bacterial communities. The DHA of soil under the combined stresses was lower than that of the control and that under the single factors, and the BIOLOG^GN substrate utilizing patterns of soil treated by combinations were distinctively differentiated from the control and their corresponding single factors. From all of above, the methamidophos, copper, especially their combinations had the clearly potential ecotoxicological risks to influence the natural soil microbial ecological system by changing the structure, richness, and the functional characteristics of microbial community.     

氧化葡萄糖酸杆菌酶学和分子生物学研究

对氧化葡萄糖酸杆菌初级代谢途径中的关键酶及分子生物学研究做了系统的评述 ,展望了分子技术改造氧化葡萄糖酸杆菌和优化 2 KGA代谢途径的可能。     

基于荧光染料Cy5.5标记的引物延伸法鉴定细菌RNA剪切加工位点

【背景】传统引物延伸法常与放射性标记结合使用,但由于放射性元素半衰期短、危害人体健康及严格的操作要求等限制了其在一般实验室的广泛使用。因此,开发新型非放射性引物延伸法成为当前研究的热点。【目的】建立非放射性的引物延伸法,并利用该方法实现对细菌RNA剪切加工位点的鉴定。【方法】将包含RNA剪切位点的序列克隆至双荧光报告系统,然后利用荧光染料Cy5.5标记的特异性靶向mcherry寡核苷酸引物进行延伸,并辅以Northern blotting、RT-qPCR等技术,确定RNA剪切加工的精确位点。【结果】鉴定了来自解纤维梭菌cip-cel mRNA中的2个剪切加工位点均位于颈环结构下游的单链区域,且在剪切位点附近未发现保守序列。【结论】基于Cy5.5标记的引物延伸法能够精确鉴定RNA剪切加工位点,且与传统的放射性引物延伸法相比,该方法具有更高的安全性和更快的检测速度,能够满足一般实验室的实验要求。     

针叶小爪螨的滞育研究

本文研究针叶小爪螨Oligonychus ununguis(Jacobi)滞育卯出现的规律,分析了光照、温度、寄主营养等环境因素与滞育的关系和滞育卵解除的条件。结果表明,针叶小爪螨以滞育卵在枝条上越冬,其产出盛期在8月上、中旬。光周期感应螨态为幼螨至第二若螨的连续两个螨态;其临界光照时间为13h/d。在相同条件下,低温(20℃)促进滞育。长日照下,螨口密度大,寄主营养恶化,亦能促进滞育的形成。滞育卵在0℃下放置1 20天,5℃-10℃下放置100天后解除滞育。     

Tat Peptide-Mediated Soluble Expression of the Membrane Protein LSECtin-CRD in Escherichia coli

The human liver and lymph node sinusoidal endothelial cell C-type lectin (hLSECtin), a type II integral membrane protein, containing a Ca²⁺-dependent carbohydrate recognition domain (CRD), has a well-established biological activity, yet its three-dimensional structure is unknown due to low expression yields and aggregation into inclusion bodies. Previous study has demonstrated that the HIV-1 virus-encoded Tat peptide (‘YGRKKRRQRRR’) can increase the yields and the solubility of heterologous proteins. However, whether the Tat peptide could promote the high-yield and soluble expression of membrane proteins in Escherichia coli is not known. Therefore, the prokaryotic expression vector pET28b-Tat-hLSECtin-CRD (using pET28b and pET28b-hLSECtin-CRD as controls) was constructed, and transformed into E. coli BL21 (DE3) cells and induced with isopropyl-β-d-thiogalactoside (IPTG) followed with identifying by SDS-PAGE and Western blot. Subsequently, the bacterial subcellular structure, in which overexpressed the heterologous proteins Tat-hLSECtin-CRD and Tat-free hLSECtin-CRD, was analyzed by transmission electron microscope (TEM) respectively, and the mannose-binding activity of Tat-hLSECtin-CRD was also determined. Expectedly, the solubility of Tat-LSECtin-CRD significantly increased compared to Tat-free LSECtin-CRD (**p < 0.01) with prolonged time, and the Tat-LSECtin-CRD had a significant mannose-binding activity. The subcellular structure analysis indicated that the bacterial cells overexpressed Tat-hLSECtin-CRD exhibited denser region compared with controls, while dot denser region aggregated in the two ends of bacterial cells overexpressed Tat-free hLSECtin-CRD. This study provided a novel method for improving the soluble expression of membrane proteins in prokaryotic systems by fusion with the Tat peptide, which may be potentially expanded to the expression of other membrane proteins.