草酸青霉可重复利用筛选标记5'氟乳清酸的构建

草酸青霉是纤维素酶高产真菌,也是科学研究的重要真菌之一.借助基因工程技术手段对工业真菌进行分子改造,可以有效提高菌株在工业生产中的经济效益,对工业菌株进行基因改造需要大量的筛选标记,目前草酸青霉中可用的筛选标记较少,因此需要构建一个草酸青霉可重复利用筛选标记转化系统.本研究构建以乳清酸核苷-5'-磷酸脱羧酶基因(pyrG)作为选择标记,以草酸青霉pyrG缺失菌株为宿主菌构建转化系统.首先,构建了含有草酸青霉pyrG表达框及其终止子TT1重复序列的可重复利用pyrG筛选标记.然后,以pyrG筛选标记作为选择标记,以草酸青霉pyrG缺失菌株Δku70ΔpyrG(pyrG∷kan)为受体菌株,敲除Δku70ΔpyrG(pyrG∷kan)菌株中的kan基因,获得菌株Δku70Δkan(kan∷R-pyrG).最后,利用筛选标记中的pyrG终止子重复序列发生自我剪切,利用氟乳清酸(5-FOA)的筛选,获得pyrG缺失,kan抗性基因缺失菌株Δku70ΔR-pyrG.本研究成功建立以pyrG为筛选标记,草酸青霉pyrG缺失菌株为受体菌的可重复利用筛选系统.     

中国人胎盘组织ABCB1基因多态性与P-gp表达水平的相关性

为了探讨中国人群胎盘组织中ABCB1基因单核苷酸多态性与其mRNA及蛋白表达水平的相关性,为妊娠期安全用药提供理论依据,本研究选取2016年6月至2018年5月于昆明医科大学第一附属医院正常分娩的200例中国汉族妇女的胎盘组织为研究对象,剪取母体侧胎盘组织,用于DNA、RNA和蛋白质的提取.通过测序确定ABCB1基因rs1045642、rs1128503及rs2032582多态性位点的基因型;采用实时荧光定量PCR和Western blot法测定组织中P-gp的mRNA和蛋白表达水平;对ABCB1基因多态性与P-gp的mRNA及蛋白表达水平的相关性进行分析.研究发现,对于rs1045642、rs1128503及rs2032582多态性位点,各基因型之间P-gp的mRNA及蛋白表达水平差异均无统计学意义(F=0.813,0.219,P＞0.05;F=0.132,0.086,P＞0.05;F=1.617,0.794,P＞0.05).本研究表明ABCB1 rs1045642、rs1128503及rs2032582基因多态性与中国汉族人群胎盘组织P-gp mRNA及蛋白表达水平可能无关.     

草菇低温胁迫初期应答关键通路的探究

本研究利用表达谱芯片技术解析了低温胁迫初期（0-120min）不同低温处理时间草菇转录组水平的变化。芯片结果分析发现在低温诱导过程中,低温处理20min内变化基因的功能为单加氧酶、氧化还原酶和细胞内的氧化还原。低温处理40min时表达差异基因具有结合RNA的功能。80min的时间段内,基因的功能比较集中于核酸物质和能量的代谢。CYP450的代谢途径在20-40min、60-80min和100-120min低温处理的菌丝体中均有显著性变化,表明这一通路在草菇低温处理过程中表现活跃。基因表达趋势分析发现34个表达差异基因富集到显著表达趋势模型,基因共表达网络分析发现VVO_04066位于网络的核心地位,推测VVO_04066通过提高磷酸肌醇特异性磷脂酶C（PI-PLC）的胞内水平,促进糖蛋白和几丁质的合成,从而完成草菇细胞的低温胁迫应答。     

Structure of plant photosystem I−light harvesting complex I supercomplex at 2.4 Å resolution

Photosystem I (PSI) is one of the two photosystems in photosynthesis, and performs a series of electron transfer reactions leading to the reduction of ferredoxin. In higher plants, PSI is surrounded by four light-harvesting complex I (LHCI) subunits, which harvest and transfer energy efficiently to the PSI core. The crystal structure of PSI-LHCI supercomplex has been analyzed up to 2.6 Å resolution, providing much information on the arrangement of proteins and cofactors in this complicated supercomplex. Here we have optimized crystallization conditions, and analyzed the crystal structure of PSI-LHCI at 2.4 Å resolution. Our structure showed some shift of the LHCI, especially the Lhca4 subunit, away from the PSI core, suggesting the indirect connection and inefficiency of energy transfer from this Lhca subunit to the PSI core. We identified five new lipids in the structure, most of them are located in the gap region between the Lhca subunits and the PSI core. These lipid molecules may play important roles in binding of the Lhca subunits to the core, as well as in the assembly of the supercomplex. The present results thus provide novel information for the elucidation of the mechanisms for the light-energy harvesting, transfer and assembly of this supercomplex.     

美洲大蠊乙醇提取物对香烟引起小鼠肺脏氧化应激的干预效果研究

美洲大蠊提取物具有显著的抗肿瘤、抗菌和抗肝纤维化等作用,但是,其对肺脏抗氧化应激效果的研究未见报道,本文主要探讨美洲大蠊乙醇提取物对熏烟小鼠血液和肺脏抗氧化应激的效果.选用雌性KM小鼠28只,随机分为对照组、熏烟组、低剂量和高剂量美洲大蠊醇提物组各7只,除对照组灌胃生理盐水外,其他各组每天中午和下午采用烟熏处理,持续31 d,其中第17天开始高剂量和低剂量组每天晚上灌胃美洲大蠊醇提物,实验结束后计算肺脏系数,检测血浆和肺脏组织匀浆中MDA、SOD、CAT、NO、GSH-Px和肺脏SOD、GSH-PX、CAT基因mRNA表达的水平以及肺脏组织病理结构的变化.结果表明:各组间肺脏系数和血浆、肺脏中总SOD和CAT的活力差异无统计学意义(P>0.05),但是与对照组相比,熏烟组显著升高了血浆和肺脏中NO以及肺脏中MDA的含量,显著降低了血浆中GSH-PX的活力和肺脏中该基因表达,差异具有统计学意义(P<0.05);而与熏烟组相比,低剂量和高剂量美洲大螊醇提物组均显著降低了血浆和肺脏中NO和MDA的水平,显著升高了肺脏GSH-PX的活力及其基因表达,高剂量组还显著升高了血浆中GSH-PX的活力,差异均具有统计学意义(P<0.05);高剂量美洲大蠊醇提物还对香烟造成的肺组织损害具有缓解作用.研究结果表明美洲大蠊乙醇提取物对熏烟小鼠血液和肺组织具有较好的抗氧化应激功能.     

家蝇Phormicin A多克隆抗体的制备及效价检测

家蝇Phormicin作为防御素家族的成员,是一类具有广普抗菌活性的抗菌肽.本研究采用原核表达法表达并纯化获得了家蝇PhormicinA蛋白.将家蝇Phormicin A原核表达蛋白与完全佐剂和不完全佐剂乳化混匀,先后免疫新西兰白兔获得其多克隆抗体.通过原核表达蛋白中和吸附实验,以及Western blot实验验证了抗体的特异性.进一步用金黄色葡萄球菌刺激家蝇三龄幼虫样品,进行内源性验证并测定抗体的效价.结果 显示,该多克隆抗体既可以识别家蝇Phormicin A原核表达蛋白,也可以识别金黄色葡萄球菌刺激家蝇三龄幼虫产生的内源性Phormicin A蛋白.本研究为进一步探索Phormicin在家蝇天然免疫和防御中的机制等后续工作打下基础.     

A practical guide to amplicon and metagenomic analysis of microbiome data

Advances in high-throughput sequencing(HTS)have fostered rapid developments in the field of microbiome research,and massive microbiome datasets are now being generated.However,the diversity of software tools and the complexity of analysis pipelines make it difficult to access this field.Here,we systematically summarize the advantages and limitations of micro-biome methods.Then,we recommend specific pipelines for amplicon and metagenomic analyses,and describe commonly-used software and databases,to help researchers select the appropriate tools.Furthermore,we introduce statistical and visualization methods suit-able for microbiome analysis,including alpha-and beta-diversity,taxonomic composition,difference compar-isons,correlation,networks,machine learning,evolu-tion,source tracing,and common visualization styles to help researchers make informed choices.Finally,a step-by-step reproducible analysis guide is introduced.We hope this review will allow researchers to carry out data analysis more effectively and to quickly select the appropriate tools in order to efficiently mine the bio-logical significance behind the data.     

Is oral microbiome of children able to maintain resistance and functional stability in response to short-term interference of ingesta?

Dear Editor, A series of studies had focused on the ecological stability of human microbiome (Lozupone et al.,2012;Faith et al.,2013;Moya and Ferrer,2016).Despite the continuous perturbation and the highly personalized composition within the human microbiome (Human Microbiome Project,2012),healthy adults stably maintain their microbial communities in terms of space and time (Faith et al.,2013;Moya and Ferrer,2016;Oh et al.,2016).This stability is proved to be critical for the well-being of human body (Lozupone et al.,2012).On the contrary,major shifts in microbial community composition are often related to diseases (Lynch and Pedersen,2016).     

A C6/C5 co‐fermenting Saccharomyces cerevisiae strain with the alleviation of antagonism between xylose utilization and robustness

During second‐generation bioethanol production from lignocellulosic biomass, the desired traits for fermenting microorganisms, such as Saccharomyces cerevisiae, are high xylose utilization and high robustness to inhibitors in lignocellulosic hydrolysates. However, as observed previously, these two traits easily showed the antagonism, one rising and the other falling, in the C6/C5 co‐fermenting S. cerevisiae strain. In this study, LF1 obtained in our previous study is an engineered budding yeast strain with a superior co‐fermentation capacity of glucose and xylose, and was then mutated by atmospheric and room temperature plasma (ARTP) mutagenesis to improve its robustness. The ARTP‐treated cells were grown in 50% (v/v) leachate from lignocellulose pretreatment with high inhibitors content for adaptive evolution. After 30 days, the generated mutant LF1‐6 showed significantly enhanced tolerance, with a six‐fold increase in cell density in the above leachate. Unfortunately, its xylose utilization dropped markedly, indicating the recurrence of the negative correlation between xylose utilization and robustness. To alleviate this antagonism, LF1‐6 cells were iteratively mutated with ARTP mutagenesis and then anaerobically grown using xylose as the sole carbon source, and xylose utilization was restored in the resulting strain 6M‐15. 6M‐15 also exhibited increased co‐fermentation performance of xylose and glucose with the highest ethanol productivity reported to date (0.525 g g^?1 h^?1) in high‐level mixed sugars (80 g L^?1 glucose and 40 g L^?1 xylose) with no inhibitors. Meanwhile, its fermentation time was shortened by 8 h compared to that of LF1. During the fermentation of non‐detoxified lignocellulosic hydrolysate with high inhibitor concentrations at pH ~3.5, 6M‐15 can efficiently convert glucose and xylose with an ethanol yield of 0.43 g g^?1. 6M‐15 is also regarded as a potential chassis cell for further design of a customized strain suitable for production of second‐generation bioethanol or other high value‐added products from lignocellulosic biomass.     

Allelopathic effects of the revegetation species Juniperus sabina L. in semiarid areas of China

Landscape and Ecological Engineering - Juniperus sabina L. is a native evergreen conifer of the Mu Us Sandy Land. It has prostrating stems that can prevent sand shifting and is an important...