大肠杆菌FC40系统静止期突变中的F因子转移

本研究采用大肠杆菌GM133 rifr细胞和营养收集细胞HB214 strr进行适应性突变实验。在混合30min和2d 后添加链霉素杀死GM133基因型细胞,继续培养5d后,在选择平板上出现了一定数量的lac+strr基因型回复突变菌落。根据这些突变菌落的数量,估计在lac+突变产生之前,GM133和HB214细胞之间的接合频率分别为0.07%和7.47%。在培养了7d的选择平板上添加含链霉素的M9选择培养基,2d 后也观察到大量发生lac+突变但没有形成肉眼可见菌落的营养收集细胞。此外,在lac+突变发生后,也有F因子从GM133细胞转移进入HB214细胞。这些事实表明,在FC40系统的适应性突变实验中发生了真正的F因子转移。 Abstract:The experiment of adaptive mutation was performed by using Escherichia coli GM133 rifr as test cells and HB214 strr as scavenger cells.Transfer frequency between GM133 and HB214 was estimated,based on the number of revertants appeared on the selective plates when GM133 were killed by addition of M9 selective medium containing 100μg/mL of streptomycin at different time.After 30 minutes the cells of GM133 and HB214 were mixed,the estimated transfer frequency was about 0.07%,and two days,7.47%.After selection of 7 days,some HB214 cells with F` factor from GM133 cells and lac+ mutation were observed,but these cells failed to form the colonies which can be seen by the naked-eye.It was demonstrated that actual F` factor transfer events from test cells GM133 to scavenger cells HB214 occurred during the selection.     

Ironing out the distribution of [2Fe-2S] motifs in ferrochelatases

Heme, a near ubiquitous cofactor, is synthesized by most organisms. The essential step of insertion of iron into the porphyrin macrocycle is mediated by the enzyme ferrochelatase. Several ferrochelatases have been characterized, and it has been experimentally shown that a fraction of them contain [2Fe-2S] clusters. It has been suggested that all metazoan ferrochelatases have such clusters, but among bacteria, these clusters have been most commonly identified in Actinobacteria and a few other bacteria. Despite this, the function of the [2Fe-2S] cluster remains undefined. With the large number of sequenced genomes currently available, we comprehensively assessed the distribution of putative [2Fe-2S] clusters throughout the ferrochelatase protein family. We discovered that while rare within the bacterial ferrochelatase family, this cluster is prevalent in a subset of phyla. Of note is that genomic data show that the cluster is not common in Actinobacteria, as is currently thought based on the small number of actinobacterial ferrochelatases experimentally examined. With available physiological data for each genome included, we identified a correlation between the presence of the microbial cluster and aerobic metabolism. Additionally, our analysis suggests that Firmicute ferrochelatases are the most ancient and evolutionarily preceded the Alphaproteobacterial precursor to eukaryotic mitochondria. These findings shed light on distribution and evolution of the [2Fe-2S] cluster in ferrochelatases and will aid in determining the function of the cluster in heme synthesis.     

Genetic analysis of resistance to Puccinia striiformis f.sp. tritici in cv. Aflak at adult plant stage

In order to investigate heritability and gene action for yellow rust resistance in wheat, a resistance yellow rust cultivar Aflak was crossed to susceptible cultivar Avocet‘s’. Parents, F₁, F₂ and F₃ generations were cultured according to randomised complete block design with two replications in the research station of Gharakhil, Iran. Parents and other generations were inoculated with 70E0A+ race. Traits including severity and infection type were recorded and then coefficient of infection was calculated. For this trait, generations mean and variance analysis were performed and results showed that there were significant differences among generations for coefficient of infection. Results showed that in addition to additive and dominance effects, at least one kind of epistasis interaction (additive × additive) control this trait. Although additive and dominance effects control this trait, but with attention to generations variance analysis, the results showed that additive variance had important role to control this trait.     

Signaling proteins in raft-like microdomains are essential for Ca2+ wave propagation in glial cells

The hypothesis that calcium signaling proteins segregate into lipid raft-like microdomains was tested in isolated membranes of rat oligodendrocyte progenitor (OP) cells and astrocytes using Triton X-100 solubilization and density gradient centrifugation. Western blot analysis of gradient fractions showed co-localization of caveolin-1 with proteins involved in the Ca2+ signaling cascade. These included agonist receptors, P2Y1, and M1, TRPC1, IP3R2, ryanodine receptor, as well as the G protein Galphaq and Homer. Membranes isolated from agonist-stimulated astrocytes showed an enhanced recruitment of phospholipase C (PLCbeta1), IP3R2 and protein kinase C (PKC-alpha) into lipid raft fractions. IP3R2, TRPC1 and Homer co-immunoprecipitated, suggesting protein-protein interactions. Disruption of rafts by cholesterol depletion using methyl-beta-cyclodextrin (beta-MCD) altered the distribution of caveolin-1 and GM1 to non-raft fractions with higher densities. beta-MCD-induced disruption of rafts inhibited agonist-evoked Ca2+ wave propagation in astrocytes and attenuated wave speeds. These results indicate that in glial cells, Ca2+ signaling proteins might exist in organized membrane microdomains, and these complexes may include proteins from different cellular membrane systems. Such an organization is essential for Ca2+ wave propagation.     

Effects of aging and calorie restriction on the global gene expression profiles of mouse testis and ovary

Background  

The aging of reproductive organs is not only a major social issue, but of special interest in aging research. A long-standing view of 'immortal germ line versus mortal soma' poses an important question of whether the reproductive tissues age in similar ways to the somatic tissues. As a first step to understand this phenomenon, we examine global changes in gene expression patterns by DNA microarrays in ovaries and testes of C57BL/6 mice at 1, 6, 16, and 24 months of age. In addition, we compared a group of mice on ad libitum (AL) feeding with a group on lifespan-extending 40% calorie restriction (CR).