Molecular cloning and characterization of double-stranded cDNA coding for bovine chymosin

A full-length cDNA copy of the mRNA encoding calf chymosin (also known as rennin), a proteolytic enzyme with commercial importance in the manufacture of cheese, has been cloned in an f1 bacteriophage vector. The nucleotide sequence of the cDNA was determined, and translation of that sequence into amino acids predicts that the zymogen prochymosin is actually synthesized in vivo as preprochymosin with a 16 amino acid signal peptide. In vitro translation of total poly(A)-enriched RNA from the calf fourth stomach (abomasum) and immunoprecipitation with antichymosin antiserum revealed that a form of chymosin (probably preprochymosin judging from the M_r-value) is the major in vitro translation product of RNA from that tissue. Gel-transfer hybridization of restriction endonuclease-cleaved bovine chromosomal DNA with labeled cDNA probes indicated that the two known forms of chymosin, A and B, must be products of two different alleles of a single chymosin gene.     

根癌农杆菌介导真菌遗传转化的研究及应用

根癌农杆菌介导转化法（Agrobacterium tumefaciens-mediated transformation,ATMT）具有转化效率高、遗传稳定、适用范围广等诸多优点,已成为真菌遗传转化研究中的强有力手段,在真菌基因资源开发、真菌性疾病研究和外源蛋白表达研究中发挥巨大作用。本文概述了根癌农杆菌转化法在真菌转化中的研究进展、技术优缺点、转化机制、实验方法和应用现状,着重介绍影响其转化效率的因素并对优化方法进行探讨,展望了该技术在真菌基因资源发掘、基因编辑等方面的应用前景,为今后真菌的遗传转化研究提供参考。     

黄土丘陵区人工柠条种植年限和坡位对土壤团聚体稳定性的影响

柠条(Caragana korshinskii Kom.)种植是黄土高原地区生态环境建设中重要的人工植被恢复措施。选择黄土丘陵区条带种植15、25和35年的柠条坡地,以荒草坡地为对照,运用Le Bissonnais法分析柠条种植不同年限和坡位对0—40 cm土层团聚体分布及其稳定性的影响。结果表明:长时间柠条种植对土壤团聚体稳定性的影响主要在0—20 cm土层;不同处理下土壤大团聚体(>0.25 mm)含量总体表现为柠条35年>柠条25年>柠条15年>荒草地,表明柠条种植年限增加促进了大团聚体的形成。从坡面尺度看,柠条平均重量直径整体表现为坡下>坡上>坡中;在上坡的柠条35年样地具有最大值(3.08 mm),但在下坡荒草地显著高于柠条林地(P<0.05)。基于相对消散指数和相对机械破碎指数,上中坡土壤团聚体均对消散作用和机械破碎作用较敏感,而下坡的下层土壤团聚体对破碎作用更敏感。冗余分析表明,土壤有机碳和粘粒含量与柠条平均重量直径呈显著正相关关系(P<0.05)。种植年限是影响土壤团聚体稳定性的主要因素,解释了30.4%的变异;其次是土层...     

温度诱导Targetron系统用于大肠杆菌高效基因失活

构建基于Te I3c/4c嗜热二型内含子的温度诱导Targetron基因失活系统(Thermotargetron),并应用于中温微生物基因编辑。在大肠杆菌HMS174(DE3)基因组中,选择Subunitofflagellum基因(fliC)和C4dicarboxylate orotate:H+symporter基因(dctA)为靶基因。根据Te I3c/4c DNA识别规则,在fliC和dctA基因中选择fliC489a、fliC828s、fliC1038s和dctA2a位点为基因打靶位点。使用重叠延伸PCR方法,基于pHK-TT1A质粒构建打靶载体。打靶载体转化HMS174菌株,对数期转化子培养液48℃热激1h后涂布于氯霉素抗性LB平板上。使用菌落PCR和DNA测序检测突变株并计算基因失活效率。获得突变株后,通过琼脂穿刺和碳源代谢实验,鉴定ΔfliC、ΔdctA突变株表型变化。菌落PCR测序结果表明,Te I3c/4c插入到fliC和dctA基因设计位点,且打靶效率高达100%。突变株表型验证实验表明,ΔfliC突变株运动能力显著下降,ΔdctA突变株苹果酸代谢能力缺失。综上所述,...     

Studying bona fide SARS-CoV-2 biology in a BSL-2 biosafety environment using a split-virus-genome system

<正>Dear Editor,Severe acute respiratory syndrome coronavirus 2(SARSCoV-2) was identified as the pathogen causing the coronavirus disease(COVID-19), which sometimes resulted in fatal pneumonia(Hu et al., 2021). SARS-CoV-2 is a biosafety level 3(BSL-3) pathogen, and the requirement for high containment conditions is a bottleneck for basic research on viral biology.     

Cryptic phenology in plants: Case studies,implications, and recommendations

Plant phenology—the timing of cyclic or recurrent biological events in plants—offers insight into the ecology, evolution, and seasonality of plant‐mediated ecosystem processes. Traditionally studied phenologies are readily apparent, such as flowering events, germination timing, and season‐initiating budbreak. However, a broad range of phenologies that are fundamental to the ecology and evolution of plants, and to global biogeochemical cycles and climate change predictions, have been neglected because they are “cryptic”—that is, hidden from view (e.g., root production) or difficult to distinguish and interpret based on common measurements at typical scales of examination (e.g., leaf turnover in evergreen forests). We illustrate how capturing cryptic phenology can advance scientific understanding with two case studies: wood phenology in a deciduous forest of the northeastern USA and leaf phenology in tropical evergreen forests of Amazonia. Drawing on these case studies and other literature, we argue that conceptualizing and characterizing cryptic plant phenology is needed for understanding and accurate prediction at many scales from organisms to ecosystems. We recommend avenues of empirical and modeling research to accelerate discovery of cryptic phenological patterns, to understand their causes and consequences, and to represent these processes in terrestrial biosphere models.     

硬覆盖对土壤水热传输及作物生长发育影响的试验研究

河北省盐渍区农业水资源非常紧缺 ,农业节水势在必行。覆盖保摘技术是农田节水管理的重要措施之一 ,国内外对此有过不少研究报道。但多集中在地膜、秸秆、砂砾和化学喷涂等覆盖材料上 ,这些覆盖材料在使用中有种种不理想之处 ,如地膜覆盖容易造成残膜的“白色污染” ,并且使用年限短 ,降水不易入渗 ,中耕、除草、施肥困难 ,使作物抗病力差等[1 ] ;秸秆覆盖容易使秸秆中的毒素物质与作物间发生生化它感现象 ,影响作物生长[2 ] 。化学喷涂除了不利于农事作业外 ,还可能造成土壤污染。为此 ,我们在中国科学院南皮试区盐渍土上试用了一种新型覆…     

Macrophage migration inhibitory factor (MIF) interacts with Bim and inhibits Bim-mediated apoptosis

The pro-apoptotic Bcl-2 family member Bim acts as a sensor for apoptotic stimuli and initiates apoptosis through the mitochondrial pathway. To identify novel regulators of Bim, we employed the yeast two-hybrid system and isolated the human gene encoding macrophage migration inhibitory factor (MIF), a ubiquitously expressed proinflammatory mediator that has also been implicated in cell proliferation, the cell cycle and carcinogenesis. The interaction between MIF and Bim was confirmed by both in vitro and in vivo protein interaction assays. Intriguingly, protein complexes between MIF and the three major Bim isoforms (BimEL/BimL/BimS) could be detected in HEK293 and K562 cells, especially in cells undergoing apoptosis. Moreover, exogenous expression of MIF partially inhibited Bim-induced apoptosis in HEK293 cells. SiRNA-mediated knockdown of MIF increased apoptosis in K562 cells exposed to the chemical oxidant diamide. Endogenous MIF may regulate the pro-apoptotic activity of Bim and inhibit the release of cytochrome c from mitochondria.     

Coarse-grained molecular dynamics simulation of interactions between cyclic lipopeptide Bacillomycin D and cell membranes

According to experimental studies, Bacillomycin D has strong antimicrobial activities, but the antimicrobial mechanism is still unknown. In this paper, the interaction mechanisms between this cyclic lipopeptide and three different charged cell membranes are studied via Coarse-Grained Molecular Dynamics (CG MD) simulations. A specific CG model for the cyclic lipopeptide Bacillomycin D was developed. The insertion of cyclic lipopeptide Bacillomycin D into DOPC, DOPC/DPPA and DOPC/DOTAP cell membranes was investigated. The position distribution and stability of Bacillomycin D in the three different cell membranes were analysed and compared based on density profile calculations. Additionally, we focused on the Radial Distribution Function (RDF) curves between amino acid residues with negative charges or strong hydrophobic properties and the head groups of two different cell membranes. Based on changes in the curvature of the three membranes, the cyclic lipopeptide Bacillomycin D can cause localised surface protrusions in DOPC/DOTAP membranes, inward depressions in the surface of DOPC/DPPA membranes and inhibition deformation in the surface of DOPC membranes. This study will help to further understand the antibacterial mechanism of the cyclic lipopeptide Bacillomycin D and provide a basis for the development of new antibiotics.