真核mRNA转录终止机制

真核RNA聚合酶Ⅱ催化的mRNA转录是基因表达中一个重要阶段,但是对它的终止过程却知之甚少。大量实验表明,真核mRNA转录终止涉及到RNA聚合酶Ⅱ最大亚基（Rpb1）C末端结构域和多种转录终止相关因子以及两者之间的相互作用。这些结果初步勾画了真核mRNA转录终止的一般过程。     

染色体步移技术研究进展

染色体步移技术是一种常用的克隆已知片段旁侧序列的技术.综述了近年来染色体步移技术的发展情况,介绍了结合基因组文库的染色体步移技术和基于PCR的染色体步移技术.同时总结了物理剪切法和限制性内切酶法构建亚克隆文库的优化步骤,以及连接成环PCR法、外源接头介导PCR法和半随机引物PCR法的原理,并且比较分析了他们之间的优缺点,以期对实际操作起到借鉴作用.     

The lipid-droplet proteome reveals that droplets are a protein-storage depot

BACKGROUND: Lipid droplets are ubiquitous organelles that are among the basic building blocks of eukaryotic cells. Despite central roles for cholesterol homeostasis and lipid metabolism, their function and protein composition are poorly understood. RESULTS: We purified lipid droplets from Drosophila embryos and analyzed the associated proteins by capillary LC-MS-MS. Important functional groups include enzymes involved in lipid metabolism, signaling molecules, and proteins related to membrane trafficking. Unexpectedly, histones H2A, H2Av, and H2B were present. Using biochemistry, genetics, real-time imaging, and cell biology, we confirm that roughly 50% of certain embryonic histones are physically attached to lipid droplets, a localization conserved in other fly species. Histone association with droplets starts during oogenesis and is prominent in early embryos, but it is undetectable in later stages or in cultured cells. Histones on droplets are not irreversibly trapped; quantitation of droplet histone levels and transplantation experiments suggest that histones are transferred from droplets to nuclei as development proceeds. When this maternal store of histones is unavailable because lipid droplets are mislocalized, zygotic histone production starts prematurely. CONCLUSIONS: Because we uncover a striking proteomic similarity of Drosophila droplets to mammalian lipid droplets, Drosophila likely provides a good model for understanding droplet function in general. Our analysis also reveals a new function for these organelles; the massive nature of histone association with droplets and its developmental time-course suggest that droplets sequester maternally provided proteins until they are needed. We propose that lipid droplets can serve as transient storage depots for proteins that lack appropriate binding partners in the cell. Such sequestration may provide a general cellular strategy for handling excess proteins.     

施肥措施对鳄嘴花(Clinacanthus nutans)生物量的影响

通过施肥措施对鳄嘴花[Clinacanthus nutans(Burm. f.) Lindau]生物量分配的影响研究得到,鳄嘴花各构件生物量与各构件生物量的分配比例并不总是一致。N肥处理组根茎叶以及总生物量均较K低。P肥则为处理4根茎叶以及总生物量较大。而K肥根茎生物量较大的均为处理5,叶和总生物量较大的则为处理4。有机肥的施肥效果对鳄嘴花各构件生物量的促进效果均较差。施肥配比中,各个施肥配比对根、茎生物量的增加效果均较差。叶和总生物量中,则为N1∶P1∶K1效果较好。总体来看,合理施肥对鳄嘴花生物量的增加有促进作用,对生物量的分配产生一定的影响。     

壳聚糖固定化AS1．398中性蛋白酶稳定性的研究

对以壳聚糖为载体,由戊二醛作交联剂。制备的固定化ＡＳ１．３９８中性蛋白酶的稳定性进行了研究。实验结果表明,固定化ＡＳ１．３９８中性蛋白酶对热、乙醇、尿素以及ｐＨ值的稳定性均有明显的提高。     

Fe- and S-Metabolizing Microbial Communities Dominate an AMD-Contaminated River Ecosystem and Play Important Roles in Fe and S Cycling

Indigenous Fe- and S-metabolizing bacteria play important roles both in the formation and the natural attenuation of acid mine drainage (AMD). Due to its low pH and Fe-S-rich waters, a river located in the Dabaoshan Mine area provides an ideal opportunity to study indigenous Fe- and S-metabolizing microbial communities and their roles in biogeochemical Fe and S cycling. In this work, water and sediment samples were collected from the river for physicochemical, mineralogical, and microbiological analyses. Illumina MiSeq sequencing indicated higher species richness in the sediment than in the water. Sequencing also found that Fe- and S-metabolizing bacteria were the dominant microorganisms in the heavily and moderately contaminated areas. Fe- and S-metabolizing bacteria found in the water were aerobes or facultative anaerobes, including Acidithiobacillus, Acidiphilium, Thiomonas, Gallionella, and Leptospirillum. Fe- and S-metabolizing bacteria found in the sediment belong to microaerobes, facultative anaerobes, or obligatory anaerobes, including Acidithiobacillus, Sulfobacillus, Thiomonas, Gallionella, Geobacter, Geothrix, and Clostridium. Among the dominant genera in the sediment, Geobacter and Geothrix were rarely detected in AMD-contaminated natural environments. Canonical correspondence analysis indicated that pH, S, and Fe concentration gradients were the most important factors in structuring the river microbial community. Moreover, a scheme explaining the biogeochemical Fe and S cycling is advanced in light of the Fe and S species distribution and the identified Fe- and S-metabolizing bacteria.     

IKBKE protein activates Akt independent of phosphatidylinositol 3-kinase/PDK1/mTORC2 and the pleckstrin homology domain to sustain malignant transformation

Serine/threonine kinase Akt regulates key cellular processes such as cell growth, proliferation, and survival. Activation of Akt by mitogenic factor depends on phosphatidylinositol 3-kinase (PI3K). Here, we report that IKBKE (also known as IKKε and IKKi) activates Akt through a PI3K-independent pathway. IKBKE directly phosphorylates Akt-Thr308 and Ser473 independent of the pleckstrin homology (PH) domain. IKBKE activation of Akt was not affected by inhibition of PI3K, knockdown of PDK1 or mTORC2 complex. Further, this activation could be inhibited by Akt inhibitors MK-2206 and GSK690693 but not the compounds (perifosine and triciribine) targeting the PH domain of Akt. Expression of IKBKE largely correlates with activation of Akt in breast cancer. Moreover, inhibition of Akt suppresses IKBKE oncogenic transformation. These findings indicate that IKBKE is an Akt-Thr308 and -Ser473 kinase and directly activates Akt independent of PI3K, PDK1, and mTORC2 as well as PH domain. Our data also suggest that Akt inhibitors targeting the PH domain have no effect on the tumors in which hyperactive Akt resulted from elevated IKBKE.     

Preliminary results on nematicidal activity from culture filtrates of Basidiomycetes against the pine wood nematode,Bursaphelenchus xylophilus (Aphelenchoididae)

One hundred and eighty one fungal species that were isolated from the fresh fruiting bodies collected in the Mountains of Pu Er County of Yunnan Province, China were tested on the pine wood nematode,Bursaphelenchus xylophilus in vitro. Each fungal species was grown in Czapek broth and potato dextrose broth (PDB). Fifteen filtrates fromAmauroderma austrosinense, Amauroderma macer, Filoboletus sp.,Laccaria tortilis, Lactarius gerardii, Lentinula edodes, Oudemansiella longipes, Oudemansiella mucida, Peziza sp.,Pleurotus sp.,Sinotermitomyces carnosus (two strains),Strobilomyces floccopus, Termitomyces albuminosus, Tylopilus striatulus grown on PDB were found to be pathogenic to the tested nematodes. Eleven filtrates fromAmanita junguillea, Amanita sp.,Daedalea sepiaria, Fistulina hepatica, Omphalotus olearius, Oudemansiella mucida, Peziza sp.,Pleurotus pulmatus, Ramaria sp.,Tricholoma conglobatum, Tylopilus striatulus grown on Czapek broth were also pathogenic to the nematodes. When screening for nematicidal potential of fungi, it is important to study the growth medium conditions necessary to obtain the optimal nematicidal effect as fungal filtrates growing on different liquid media showed a very inconsistent toxicity towards nematodes.     

Optimization of sulfated modification conditions of tremella polysaccharide and effects of modifiers on cellular infectivity of NDV

Based on our previous research, sulfated modification conditions of Tremella polysaccharide (TPS), the chlorosulfonic acid to pyridine (CSA-Pry) ratio, reaction temperature and time, were optimized by L₉ (3⁴) orthogonal design taking the yield and degree of sulfation (DS) of modifiers as indexes. Two TPSs, TPS_tp and TPS_70c, were modified under optimized conditions. The effects of two modifiers, sTPS_tp and sTPS_70c, on cellular infectivity of NDV were determined by MTT method taking the non-modified TPS_tp, TPS_tc and TPS_70c as controls. The results showed that the optimized modification conditions were reaction temperature of 80 °C, CSA-Pry ratio of 1:6 and reaction time of 1.5 h. Five polysaccharides at proper concentrations could significantly inhibit the infectivity of NDV to CEF. The virus inhibitory rates of sTPS_tp at 1.563 μg mL⁻¹ group were the highest and significantly higher than those of other three non-modified polysaccharide groups in three sample-adding modes. This indicated that sulfated modification could significantly improve the antiviral activity of TPS. sTPS_tp possessed the best efficacy and would be as a component of antiviral polysaccharide drug.