Construction of DNA damage response gene pprI functiondeficient and function-complementary mutants in Deinococcus radiodurans

PprI, a DNA damage response factor from the extraordinary radioresistant bacterium Deinococcus radiodurans, plays a central regulatory role in multiple DNA damage repair. In this study, a fusion DNA fragment carrying kanamycin resistance gene with the D. radiodurans groEL promoter was cloned by PCR amplification and reversely inserted into the pprl locus in the genome of the wild-type strain R1. The resultingpprl-deficient strain, designated YR1,was very sensitive to ionizing radiation. Meanwhile, the recombinant DNA fragment was cloned into the shuttle vector pRADZ3, and resulted in plasmid pRADK with kanamycin resistance in D. radiodurans. The fragments containing complete pprl gene and 3‘-terminal deletion pprIΔ were cloned into plasmid pRADK. The resulted plasmids designated pRADKpprI and pRADKpprIΔ were then transformed to YR1. Results show that YR1 carrying pRADKpprI was able to fully restore the extreme radioresistance to the same level as the wild-type D. raiodurans R1, whereas YR1 pRADKpprIΔ failed to do so. Construction of DNA repair switch PprI function-deficient and function-complementary mutants in D. radiodurans is not only useful to elucidating the relationship between domains and functions of PprI protein, but also opens the door to the further studies of the biological functions of PprI protein in vivo.     

Engineering <Emphasis Type="Italic">Deinococcus radiodurans</Emphasis> into biosensor to monitor radioactivity and genotoxicity in environment

Based on a genetically modified radioresistant bacteria Deinococcus radiodurans, we constructed a real time whole cell biosensor to monitor radioactivity and genotoxicity in highly radioactive environment. The enhanced green fluorescence protein （eGFP） was fused to the promoter of the crucial DNA damage-inducible recA gene from D. radiodurans, and the consequent DNA fragment （PrecA-egfp） carried by plasmid was introduced into D. radiodurans R1 strain to obtain the biosensor strain DRG300. This engineered strain can express eGFP protein and generate fluorescence in induction of the recA gene promoter. Based on the correlation between fluorescence intensity and protein expression level in live D. radiodurans cells, we discovered that the fluorescence induction of strain DRG300 responds in a remarkable dose-dependent manner when treated with DNA damage sources such as gamma radiation and mitomycin C. It is encouraging to find the widely detective range and high sensitivity of this reconstructed strain comparing with other whole cell biosensors in former reports. These results suggest that the strain DRG300 is a potential whole cell biosensor to construct a detective system to monitor the biological hazards of radioactive and toxic pollutants in environment in real time.     

Engineering Deinococcus radiodurans into biosensor to monitor radioactivity and genotoxicity in environment

Based on a genetically modified radioresistant bacteria Deinococcus radiodurans, we constructed a real time whole cell biosensor to monitor radioactivity and genotoxicity in highly radioactive environ-ment. The enhanced green fluorescence protein (eGFP) was fused to the promoter of the crucial DNA damage-inducible recA gene from D. radiodurans, and the consequent DNA fragment (PrecA-egfp) car-ried by plasmid was introduced into D. radiodurans R1 strain to obtain the biosensor strain DRG300. This engineered strain can express eGFP protein and generate fluorescence in induction of the recA gene promoter. Based on the correlation between fluorescence intensity and protein expression level in live D. radiodurans cells, we discovered that the fluorescence induction of strain DRG300 responds in a remarkable dose-dependent manner when treated with DNA damage sources such as gamma radiation and mitomycin C. It is encouraging to find the widely detective range and high sensitivity of this re-constructed strain comparing with other whole cell biosensors in former reports. These results suggest that the strain DRG300 is a potential whole cell biosensor to construct a detective system to monitor the biological hazards of radioactive and toxic pollutants in environment in real time.     

Deletion of an electron donor-binding subunit of the NDH-1 complex,NdhS, results in a heat-sensitive growth phenotype in Synechocystis sp. PCC 6803

In cyanobacteria and higher plants, NdhS is suggested to be an electron donor-binding subunit of NADPH dehydrogenase （NDH-1） complexes and its absence impairs NDH-l-dependent cyclic electron trans- port around photosystem I （NDH-CET）. Despite significant advances in the study of NdhS during recent years, its functional role in resisting heat stress is poorly understood. Here, our results revealed that the absence of NdhS resulted in a serious heat-sensitive growth phenotype in the uni- cellular cyanobacterium Synechocystis sp. strain PCC 6803. Furthermore, the rapid and significant increase in NDH-CET caused by heat treatment was completely abolished, and the repair of photosystem II under heat stress conditions was greatly impaired when compared to that of other photosynthetic apparatus in the thylakoid membrane. We therefore conclude that NdhS plays an important role in resistance to heat stress, possibly by stabilizing the electron input module of cyanobacterial NDH-1 complexes.     

dr1127： A novel gene of Deinococcus radiodurans responsible for oxidative stress

The functional analysis of dr1127,a novel gene in Deinococcus radiodurans was performed in this pa-per. The dr1127 gene was found occasionally in our microarray and 2-DE gel experiments. Mutation of the dr1127 gene decreased the γ-radiation and H2O2 resistance of D. radiodurans,and weakened the scavenging abilities of cell extracts for free radicals (superoxide anion,hydrogen peroxide,and hy-droxyl radical). Further oxidative damage assays demonstrated that the purified DR1127 protein of D. radiodurans could bind to double stranded DNA in vitro and protect DNA from oxidative damage in this way. These results suggest that the dr1127 gene is an important gene that can maintain γ-radiation and oxidative resistance in D. radiodurans and may take part in the oxidative stress process.     

Duplication and Combination of P-Loop Containing Nucleotide Triphosphate Hydrolases Superfamily

In a genome the set of proteins are formed by duplication and combination of domain superfamilies. P-loop containing nucleotide triphosphate （NTP） hydrolases superfamily is massively duplicated and has the most different partner superfamilies among archaea, bacteria and eukarya, Here, we study the distributions of duplication and combination of p-loop containing NTP hydrolases superfamily in 169 completed genomes. When the total number of domains in a genome is larger, duplication and combination partners of p-loop conraining NTP hydrolases are more. This phenomenon is more obvious in metazoa. The distributions of abundance and corn bination of partners relate to the functions of the protein. Those distributions in metazoa are very different from those in other kingdoms because of complexity of metazoa. Finally the relationship between duplication and combination of p-loop containing NTP hydrolases superfamily in different genomes is described. It fits a power law.     

Flaking and degradation of polyurethane coatings after 2 years of outdoor exposure in Lhasa

The exposure of a high gloss polyurethane aircraft topcoat system with polyurethane intermediate coating and epoxy primer to strong ultraviolet radiation in Lhasa, Tibet was studied by using scanning electron microscopy (SEM), atomic force microscopy (AFM), attenuated total reflectance Fourier-transform infrared (ATR-FTIR), and X-ray photoelectron spectroscopy (XPS). Micro-flakes were found to form on the topcoat surface after exposure and the size and quantity of the flakes increased with the increase of the exposure time. The magnitude of micro-bulges and the topcoat roughness increased after a long exposure to sunlight. Flakes and micro-bulges resulted in the loss of the topcoat gloss. ATR-FTIR and XPS analysis indicated that chain scissions of the polyurea C==O group and the polyurethane C―O―C and CO―NH groups occurred, and the degradation products of NH and carbonyl groups formed due to the exposure. The breaking of the chemical structure of the polyurethane resulted in a decrease in its elasticity and in the adhesion of the flakes to its surface.     

Physiological Defense Mechanism of Ligularia intermedia Against UV—B Radiation on Dongling Mountain

Liguloria intermedia growing at different altitudes were used to investigate the mechanism of ultraviolet (UV)-resistance on physiological aspects in the field. The tests compared the absorbance of the UV-absorbing compound, the content of chlorophyll a, chlorophyll b, and carotenoids, and the activities of peroxidase (POX) and superoxide dismutase (SOD) in Ligularia intermedia growing at three different altitudes on Dongling Mountain in northern China. There were no significant differences between the plants growing at 1160m and 1820m. However, all of these factors increased dramatically at 2190m. The results indicate tha tthe UV-resistance of the plants mainly depends on the mechanism of filtering the radiation and preventing the reactive oxygen species (ROS) damage produced by UV-B.     

内蒙古额吉淖尔盐碱湖嗜盐古菌的抗生素抗性及分泌的嗜盐菌素的抑菌性

本实验室从内蒙古额吉淖尔湖盐碱土样分离得到一株属于Natrinema属的嗜盐古菌。本文对其进行了抗生素抗性实验以及对其产生的嗜盐菌素进行了抑菌性研究。结果表明诺氟沙星对该菌具有较强的抑菌作用,头孢氨苄对该菌具有微弱的抑菌作用。通过测定抑菌圈的大小表明该菌株对抗生素具有一定的抗性。该菌株所生产的嗜盐菌素对蛋白酶K 较敏感,对大肠杆菌和金黄色葡萄球菌具有抑菌作用。     

中国云南地区地面紫外辐射观测

 在云南丽江(26°52′N,100°13′E)和勐腊(21°29′N,101°34′E)两地分别对以254nm和297nm为中心波长的太阳紫外辐射作了地面观测.观测时间为1999年7月～2000年5月.本文分析了两地紫外辐射的时空变化并初步讨论了与云量、臭氧总量的相关关系.研究结果表明:297nm波段的紫外辐射夏季强,冬季弱,具有明显的季节变化.丽江地区的紫外辐射在同一云量条件下高于勐腊地区,这可能是由海拔高度的差异引起.云量对两地的紫外辐射变化有强烈的影响,而当剔除云量、季节和随机变化等影响因素后,紫外辐射和臭氧总量呈现出了负的相关关系.     

紫外图像弱小目标检测与跟踪

工作在“日盲”波段的紫外信号探测,屏蔽了太阳紫外辐射这一最大背景噪声干扰源,可以有效地进行微弱紫外目标信号采集,但是仍然面临着目标/噪声难以区分、目标短暂丢失等问题.文中对微弱紫外目标的成像特性进行详细分析,根据紫外目标成像特征提出了一种紫外图像弱小目标检测跟踪方案.该方案首先采用基于连通域的聚类方法对紫外图像中的样本点进行聚类,标识出前景中的目标块和噪声块,然后利用真实目标的轨迹特性,结合卡尔曼滤波预测目标的运动参数,从而实现对目标的检测跟踪.仿真实验证明了该方案的可行性及有效性.