一株微生物采油菌的16S rDNA鉴定和绿色荧光蛋白分子标记

筛选到一株高效生产多糖的YX菌,油田先导试验证明该菌可以用于微生物调剖提高原油采收率（Microbial Enhanced Oil Recovery,MEOR）.经16SrDNA分析鉴定,YX菌属于肠内杆菌属（Enterobacter.）,为跟踪监测其动态信息,构建表达绿色荧光蛋白（green fluorescent protein,GFP）的重组质粒pET-30a（＋）-EGFP,利用CaCl2法将其转化至YX菌,利用荧光显微镜可以观察到重组YX（GFP）工程菌的绿色荧光.结果表明YX菌可以被绿色荧光蛋白基因标记,用于微生物采油研究.     

Electrophysiological assessment of primary cortical neurons genetically engineered using iron oxide nanoparticles

The development of safe technologies to genetically modify neurons is of great interest in regenerative neurology,for both translational and basic science applications.Such approaches have conventionally been heavily reliant on viral transduction methods,which have safety and production limitations.Magnetofection (magnet-assisted gene transfer using iron oxide nanoparticles as vectors) has emerged as a highly promising non-viral alternative for safe and reproducible genetic modification of neurons.Despite the high potential of this technology,there is an important gap in our knowledge of the safety of this approach,namely,whether it alters neuronal function in adverse ways,such as by altering neuronal excitability and signaling.We have investigated the effects of magnetofection in primary cortical neurons by examining neuronal excitability using the whole cell patch clamp technique.We found no evidence that magnetofection alters the voltage-dependent sodium and potassium ionic currents that underpin excitability.Our study provides important new data supporting magnetofection as a safe technology for bioengineering of neuronal cell populations.     

Fluorescence lifetime imaging of coral fluorescent proteins

Corals, like many other coelenterates, contain fluorescent pigments that show considerable homology with the well known green fluorescent protein of the jellyfish Aequoria. In corals, unlike jellyfish, multiple proteins are present and the range of excitations and emissions suggest the possibility of energy transfer. The occurrence of F?rster resonant energy transfer (FRET) between fluorescent proteins in corals has already been reported and time-resolved spectra have shown the effect on fluorescent lifetime, but without any spatial resolution. Lifetime confocal microscopy offers lower time resolution but excellent spatial resolution. Lifetimes of the isolated A. millepora pigments amilFP490, amilFP504, and amilFP593 (names indicate emission peaks) were 2.8, 2.9, and 2.9 ns, respectively. In the coral sample, imaging the entire emission spectrum from 420 nm, the mean lifetime was reduced to 1.5 ns, implying that FRET was occurring. Looking just at the fluorescence from FRET donors the lifetime was even shorter, at 1.3 ns, supporting this interpretation. In contrast, no reduction in lifetime is seen in the coral Euphyllia ancora, where the pigment distribution also suggests that the pigments are unlikely to be involved in photoprotection. This study set out to determine the extent of FRET between pigments in two corals, Acropora millepora and Euphyllia, ancora which differ in the arrangement of their pigments and hence possibly in pigment function.     

Photoinduced electron transfer mechanism between green fluorescent protein molecules and metal oxide nanoparticles

Green fluorescent protein (GFP) molecules are attached to titanium dioxide and cadmium oxide nanoparticles via sol–gel method and fluorescence dynamics of such a protein–metal oxide assembly is investigated with a conventional time correlated single photon counting technique. As compared to free fluorescent protein molecules, time-resolved experiments show that the fluorescence lifetime of GFP molecules bound to these metal oxide nanoparticles gets shortened dramatically. Such a decrease in the lifetime is measured to be 22 and 43 percent for cadmium oxide and titanium dioxide respectively, which is due to photoinduced electron transfer mechanism caused by the interaction of GFP molecules (donor) and metal oxide nanoparticles (acceptor). Our results yield electron transfer rates of 3.139×10⁸ s⁻¹ and 1.182×10⁸ s⁻¹ from the GFP molecules to titanium dioxide and cadmium oxide nanoparticles, respectively. The electron transfer rates show a marked decrease with increasing driving force energy. This effect represents a clear example of the Marcus inverted region electron transfer process.     

Dendrimer as nanocarrier for drug delivery

Dendrimers are novel three dimensional, hyperbranched globular nanopolymeric architectures. Attractive features like nanoscopic size, narrow polydispersity index, excellent control over molecular structure, availability of multiple functional groups at the periphery and cavities in the interior distinguish them amongst the available polymers. Applications of dendrimers in a large variety of fields have been explored. Drug delivery scientists are especially enthusiastic about possible utility of dendrimers as drug delivery tool. Terminal functionalities provide a platform for conjugation of the drug and targeting moieties. In addition, these peripheral functional groups can be employed to tailor-make the properties of dendrimers, enhancing their versatility. The present review highlights the contribution of dendrimers in the field of nanotechnology with intent to aid the researchers in exploring dendrimers in the field of drug delivery.     

Circular Dichroism and Circular Polarized Luminescence from a Green Fluorescent Protein—Initial Research for Chiroptical Emission of Biological Materials

Optical properties of a green fluorescent protein (GFP) are examined using optical absorption, circular dichroism (CD), and circular polarized luminescence (CPL) spectroscopies. The GFP has chiroptical activity and exhibits green circular polarized emission, although the g _em-factor is small. Poly(vinyl alcohol) (PVA)/GFP composite films are prepared to attempt long-term preservation of the GFP emission activity. After five years, the transparent PVA/GFP composite film still exhibits stable fluorescence that appears similar to the emission from the Aequorea jellyfish.     

Automation for High-throughput Identification and Picking of GFP Expressing Colonies

Green fluorescent protein (GFP) has many applications as a marker in living cells, and has become widely used as a reporter gene in microbial, plant and animal cells. Screening microbial colonies for GFP expression enables various types of assays (e.g. for secretory mutations). However, this is laborious, non-quantitative and potentially hazardous to the operator (due to UV illumination) when performed manually. In order to address this the GloPix robot was developed. The imaging system discriminates between colonies based on the level of fluorescence activity and the picking function automatically transfers cells to microplate wells. Measuring fluorescent activity allows quantitation of fluorescent tag concentration/expression.     

New modules for PCR-based gene targeting in Candida albicans: rapid and efficient gene targeting using 100 bp of flanking homology region

The use of PCR-based techniques for directed gene alterations has become a standard tool in Saccharomyces cerevisiae. In our efforts to increase the speed of functional analysis of Candida albicans genes, we constructed a modular system of plasmid vectors and successfully applied PCR-amplified functional analysis (FA)-cassettes in the transformation of C. albicans. These cassettes facilitate: (a) gene disruptions; (b) tagging of 3'-ends of genes with green fluorescent protein (GFP); and (c) replacements of endogenous promoters to achieve regulated expression. The modules consists of a core of three selectable marker genes, CaURA3, CaHIS1 and CaARG4. Modules for C-terminal GFP-tagging were generated by adding GFP-sequences flanked at the 5'-end by a (Gly-Ala)3-linker and at the 3'-end by the S. cerevisiae URA3-terminator to these selection markers. Promoter exchange modules consist of the respective marker genes followed by the regulatable CaMAL2 or CaMET3 promoters at their 3'-ends. In order to ensure a reliably high rate of homologous gene targeting, the flanking homology regions required a size of 100 bp of gene-specific sequences, which were provided with the oligonucleotide primers. The use of shorter flanking homology regions produced unsatisfactory results with C. albicans strain BWP17. With these new modules only a minimal set of primers is required to achieve the functional analysis of C. albicans genes and, therefore, provides a basic tool to increase the number of functionally characterized C. albicans genes of this human pathogen in the near future.     

MSTP技术发展与应用

MSTP多业务传送平台基于SDH技术，同时实现TDM、ATM、以太网等业务的接入、处理和传送，并提供统一的网管，它具有SDH,ATM以及以太网／IP处理功能，通过将多业务汇聚并高效适配的方式可以实现多种业务的综合传送。论文首先回顾了MSTP发展、接着报告了MSTP相关的主要技术，从中可以大致了解MSTP的原理和其今后相关技术发展的趋势。