Broadening the versatility of lentiviral vectors as a tool in nucleic acid research via genetic code expansion

With the aim of broadening the versatility of lentiviral vectors as a tool in nucleic acid research, we expanded the genetic code in the propagation of lentiviral vectors for site-specific incorporation of chemical moieties with unique properties. Through systematic exploration of the structure–function relationship of lentiviral VSVg envelope by site-specific mutagenesis and incorporation of residues displaying azide- and diazirine-moieties, the modifiable sites on the vector surface were identified, with most at the PH domain that neither affects the expression of envelope protein nor propagation or infectivity of the progeny virus. Furthermore, via the incorporation of such chemical moieties, a variety of fluorescence probes, ligands, PEG and other functional molecules are conjugated, orthogonally and stoichiometrically, to the lentiviral vector. Using this methodology, a facile platform is established that is useful for tracking virus movement, targeting gene delivery and detecting virus–host interactions. This study may provide a new direction for rational design of lentiviral vectors, with significant impact on both basic research and therapeutic applications.     

噪音环境下花臭蛙求偶鸣声特征分析

2012年7月份,在黄山浮溪地区利用超声录音设备录制并分析了繁殖季节雄性花臭蛙(Odorrana schmackeri)个体的求偶鸣声。观察发现花臭蛙繁殖活动主要集中在7月中旬,繁殖高峰期活动无昼夜规律,全天均可见求偶鸣叫及抱对产卵等行为,并且多在浅水滩处活动。花臭蛙鸣声根据音节数和声谱特征可分为4种类型:即单音节音、婴儿音、双音节音和多音节断奏音,其中,单音节音、双音节音和婴儿音较为常见。利用Selena软件给出4种声音的语图以及各自对应的能谱图,利用Sound Analysis pro v1.2对单音节音、双音节音和婴儿音的鸣声特征参数进行定量分析,分析的声音参数包括鸣叫持续时间、音节数、音节持续时间、音节间隔、主频、脉冲率等。结果表明,花臭蛙鸣声的主频范围为1.8~4.5 k Hz(n=65)。鸣声不包括超声组分,主频峰值(3.1±0.7)k Hz,与前人电生理实验所得花臭蛙听觉敏感峰值一致,说明花臭蛙主要在这一频段进行通讯。对3种常见鸣叫音声音参数的单因素方差分析结果表明,双音节音与单音节音和婴儿音在声音持续时间上存在显著性差异(P0.01),双音节音和单音节音在第二谐波声强上也具有显著性差异(P=0.01)。花臭蛙的双音节音在3种常见鸣叫音中具有最长的持续时间,为(99.5±8.4)ms,故推测,双音节音为花臭蛙繁殖期主要求偶鸣声,并通过其鸣声时长的变化来体现自身品质的好坏。     

Absence of mutation in miR-34a gene in a Chinese longevity population

DEAR EDITOR:Centenarians are a typical longevity model characterized by delayed onset of morbidity in age-related diseases such as cancer,cardiovascular disease,dementia,and stroke(Andersen et al,2012).Though there may be a number of underlying mechanisms behind this longevity,curiously it seems that the survival advantage persists in their offspring(Terry et al,2003),suggesting a potentially important role for genetic factors.Previous studies suggested that the heritability of human longevity may be~25%(Herskind et al,1996;Mcgue et al,1993),whicih is consistent with other studies on model     

<Emphasis Type="Italic">Paradevosia shaoguanensis</Emphasis> gen. nov., sp. nov., Isolated from a Coking Wastewater

A Gram staining negative, rod-shaped, aerobic bacterial strain J5-3^T with a single polar flagellum was isolated from coking wastewater collected from Shaoguan, Guangdong, China. It was motile and capable of optimal growth at pH 6–8, 30 °C, and 0–2 % (w/v) NaCl. Its predominant fatty acids were 11-methyl C_18:1 ω7c (29.2 %), C_16:0 (20.6 %), C_19:0 cyclo ω8c (18.2 %), C_18:0 (11.0 %), and C_18:1 ω7c/C_18:1 ω6c (10.9 %) when grown on trypticase soy agar. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, two unknown glycolipids (GL1, GL2), and two unknown phospholipid (PL1, PL2). The predominant ubiquinone was Q-10, and the genome DNA G+C content was 61.7 mol %. Phylogenetic analysis based on the 16S rRNA gene sequences indicated that strain J5-3^T belonged to the family Hyphomicrobiaceae in Alphaproteobacteria. It shared the 16S rRNA gene sequence similarities of 93.8–96.1 % with the genus Devosia, 94.5–94.8 % with the genus Pelagibacterium, and <92.0 % with all the other type strains in family Hyphomicrobiaceae. It can be distinguished from the closest phylogenetic neighbors based on several phenotypic and genotypic features, including α-galactosidase activity, tetracycline susceptibility, major fatty acid composition, polar lipid profile, DNA gyrase B subunit (gyrB) gene sequence, and random-amplified polymorphic DNA profile. Therefore, we consider strain J5-3^T to represent a novel species of a novel genus within the family Hyphomicrobiaceae, for which the name Paradevosia shaoguanensis gen. nov., sp. nov. is proposed. The type strain of Paradevosia shaoguanensis is J5-3^T (=CGMCC 1.12430^T =LMG 27409^T).     

拟南芥HyPRP蛋白AZI1在大肠杆菌中的表达及其抗真菌活性分析

AZI1(AZELAIC ACID INDUCED 1)基因位于拟南芥4号染色体上,编码产物是脂质转移蛋白(lipid transfer protein, LTP)家族的一个成员。该基因在系统获得抗性(systemic acquired resistance, SAR)中具有重要功能,名称来自它可以被壬二酸(azelaic acid, AzA)诱导。已有的研究结果显示,在拟南芥中由AzA和甘油-3-磷酸(glycerol-3-phosphate, G3P)诱导的SAR反应需要AZI1和DIR1,这两个脂质转移蛋白有助于G3P的积累。为了确定AZI1蛋白是否具有抗真菌活性,本工作构建了原核表达载体pET28a-AZI1,利用大肠杆菌BL21(DE3)受体细胞制备了没有信号肽的AZI1重组蛋白。Western免疫印迹分析发现通过半乳糖苷类似物IPTG诱导表达的AZI1重组蛋白主要以包涵体的形式存在。体外抑菌实验以及激光共聚焦显微观察结果表明,用镍离子亲和层析树脂纯化的AZI1重组蛋白对灰霉菌、赤霉菌、棉花枯萎病菌和酿酒酵母细胞的生长/分裂均具有抑制作用。