细菌与噬菌体的相互抵抗作用机制

噬菌体是地球生物圈里数量最多、存在最久的个体之一,也是应对抗生素耐药细菌感染的极具特点的候选制剂之一.本文分别以细菌和噬菌体的视角,从阻止噬菌体吸附、超感染排除、限制修饰系统、CRISPR-Cas、流产感染等方面综述了细菌抗噬菌体的机制以及噬菌体针对细菌抗性机制的应变.     

细菌抗噬菌体的免疫功能及其研究进展

在抵御噬菌体侵袭的生存竞争中,细菌进化出了完善的免疫系统,包括先天免疫和适应性免疫、被动免疫和主动免疫等多种免疫方式。本综述系统地分析了细菌的多样化免疫防御功能,包括细菌通过调控噬菌体受体基因表达、分泌胞外基质遮蔽及利用受体竞争性抑制剂等多样化的策略抑制噬菌体吸附;借助超感染排斥(superinfection exclusion, Sie)系统阻止噬菌体DNA进入细胞内部;利用限制修饰(restriction-modification, RM)系统和CRISPR-Cas系统等阻断噬菌体DNA的复制和表达。如果噬菌体成功突破RM系统和CRISPR-Cas系统这两道防线,细菌可通过毒素-抗毒素(toxin-antitoxin, TA)系统和Rex系统等流产感染诱导宿主细胞死亡来遏制噬菌体的进一步扩散。同时,本研究分析了不同抗噬菌体机制间的协调作用和相互关系,并对未来可能的研究方向进行了展望。     

噬菌体基因组编辑技术及其研究进展

阐述了噬菌体的基因整合、缺失、点突变等内容,并汇总噬菌体的基因组编辑方法,旨在为噬菌体的进一步应用和研究作铺垫.     

细菌的噬菌体感染抗性机制

噬菌体广泛存在于生态环境中。细菌在与噬菌体长期的共进化过程中,衍化出了多种针对噬茵体感染的抗性机制。我们从宿主菌的抑制吸附、阻止噬菌体DNA注入、切断噬菌体DNA和影响其功能及流产感染等方面,对宿主菌抵抗噬菌体感染的机制进行了综述。     

噬菌体与细菌相互作用的分子机制研究进展

噬菌体与细菌是自然界中存在最广泛的两类微生物,两者在群体水平、个体水平以及分子水平上均存在复杂的相互作用关系.细菌能够影响溶原性噬菌体的溶原-裂解决策,而被噬菌体感染的细菌基因表达谱也会受到噬菌体影响,使宿主菌的代谢、应激、抵抗力、毒性等多种性状发生改变.现从细菌和噬菌体两者的角度,分别综述细菌抵抗噬菌体感染以及噬菌体...     

细菌毒素-抗毒素系统在噬菌体流产感染中的作用

细菌常受到数量众多的噬菌体感染,宿主细菌在和噬菌体竞赛中进化出多样化的分子策略,流产感染(abortive infection,Abi)是其中之一。毒素-抗毒素系统(toxin-antitoxin system,TA)会在细菌受到压力胁迫时表达并介导细菌的低代谢甚至休眠,还能直接减少子代噬菌体形成。此外,部分毒素序列和结构与Cas蛋白高度同源,噬菌体甚至会编码抗毒素类似物来阻遏对应毒素的活性。这表明流产感染中细菌死亡过程导致的噬菌体感染失败与TA功能高度重合,TA可能是噬菌体侵染宿主的主要阻力和防御力量之一。文中基于TA系统的分类和功能,对参与噬菌体流产感染的TA系统进行了综述,并预测具有流产功能的TA系统和其在抗生素开发和疾病治疗中的应用前景。这有助于认识细菌-噬菌体相互作用,并指导噬菌体治疗和合成生物学。     

噬菌体基因工程技术与应用研究进展

抗生素耐药性已成为全球人类健康面临的重大威胁,医药、工业、农业生产以及生态等领域均受到多重耐药菌的严重威胁。多重耐药菌感染逐渐呈现高发病率、高死亡率的趋势。噬菌体可以特异性裂解多重耐药病原菌,然而由于噬菌体宿主谱狭窄、基因组中含有不利基因等因素的制约,当前只有部分噬菌体成功应用于防治多重耐药菌感染等领域。噬菌体基因工程具有可编辑、高效等优势,为拓宽噬菌体宿主谱、设计“安全、绿色、高效”的新型噬菌体提供了理论基础。综述了噬菌体基因工程技术的研究进展,以及噬菌体在临床抗耐药菌感染、农业生产和生态环境等方面的实际应用,为噬菌体的定向改造及其在各领域中的有效应用提供了理论支持和参考。     

细菌与噬菌体相互抵抗机制研究进展

噬菌体作为一种侵染细菌的病毒,能够特异性识别宿主细菌。近年来,抗生素的过度使用导致耐药细菌的出现,噬菌体有望成为对抗耐药细菌的新武器。在细菌与噬菌体长期共进化过程中,二者都演化出一系列抵御策略。本文从抑制噬菌体吸附、阻止噬菌体DNA进入、切割噬菌体基因组、流产感染以及群体感应对噬菌体的调控等方面,对细菌抵抗噬菌体的机制以及噬菌体应对细菌的策略进行了综述,同时还列举了细菌和噬菌体相互抵抗机制的检测方法,以期为噬菌体在细菌控制中的应用以及探究细菌抵抗噬菌体的机制提供理论依据。     

CRISPR-Cas系统与细菌和噬菌体的共进化

细菌在适应噬菌体攻击的过程中,进化了多种防御系统,噬菌体在细菌的选择压力下,也在不断进化反防御策略,双方的这种进化关系与发生机制一直尚不完全清楚。近年在细菌和古细菌中发现一种新的免疫防御系统,即CRISPR-Cas(clustered regularly interspaced short palindromic repeats-CRISPR-associated system)系统。在对其功能和作用机制深入研究的同时,也不断地揭示了细菌和噬菌体之间的共进化关系。为此,文章在介绍原核细胞中CRISPR-Cas系统介导的免疫机制基础上,重点综述了CRISPR系统在细菌和噬菌体进化中的作用。     

噬菌体表面呈现技术研究进展

噬菌体表面呈现技术是1985年建立的一种将外源基因表达呈现在噬菌体颗粒表面的方法,可用于建立随机多肽文库、抗体文库等。经特定配基的筛选,可获得与其特异结合的配体分子。通过改构,还可将cDNA产物表达于噬菌体颗粒的尾部构建cDNA文库。SIP技术通过将配体和配基分别与基因Ⅲ蛋白的C末端和N末端融合表达,基因Ⅲ的C-末端参与噬菌体颗粒的组装,配基与配体的结合能够重建基因Ⅲ蛋白的功能,才能形成有感染能力的噬菌体,这样就大大提高了筛选效率。     

基于CRISPR/Cas系统的噬菌体基因组编辑

对原核生物获得性免疫系统CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR- associated genes)的研究促进了新一代基因组编辑工具的产生和发展。噬菌体既是原核生物CRISPR阵列(CRISPR array)进化的原动力,又是CRISPR/Cas系统防御的对象。噬菌体功能基因组学研究的速率却落后于发现新噬菌体和测定基因组序列的速率。基于CRISPR/Cas系统的噬菌体基因组编辑,可为噬菌体功能基因组学研究提供新手段。本文评述了基于CRISPR/Cas系统编辑噬菌体基因组的几例开创性研究,并且比较了多种操作程序的异同点和优缺点。同时,进一步构建了联合使用CRISPR/Cas系统与噬菌体重组系统开展噬菌体基因组编辑的新方案,讨论了新方案的潜在局限性,并对如何选择不同方案给予了建议。     

Resistance of corynebacterial strains to infection and lysis by corynephage BFK 20

AIMS: Defence mechanisms of the corynebacterial strains against corynephage BFK 20, which causes lysis of Brevibacterium flavum CCM 251. METHODS AND RESULTS: We tested adsorption of the phage BFK 20 to the corynebacterial cell surface. We observed strong adsorption ranging from ca 79 to 93% on the cells of B. flavum ATCC strains, but only ca 76% for B. flavum CCM 251. Minor adsorption for Brevibacterium lactofermentum BLOB (ca 13%) and no adsorption for Corynebacterium glutamicum RM3 were determined. BFK 20 infection had no significant effect on growth and viability of C. glutamicum and B. lactofermentum, but significantly influenced growth and viability of B. flavum ATCC 21127, 21128 and 21474. Cell growth stopped in short time after infection but with no lysis. Brevibacterium flavum CCM 251 cell growth was arrested too and lysis occurred. The Southern hybridization confirmed the presence of significant amount of BFK 20 DNA in samples from B. flavum CCM 251 and B. flavum ATCC strains after BFK 20 infection. Only weak hybridization signal was detected for DNA from infected cells of B. lactofermentum BLOB and no signal for C. glutamicum RM3. CONCLUSIONS: Based on the above results we suggest presence of a mechanism leading to abortive infection in B. flavum ATCC 21127, 21128 and 21474. In B. lactofermentum BLOB and C. glutamicum RM3 the adsorption barrier is more likely. SIGNIFICANCE AND IMPACT OF THE STUDY: This study increases the knowledge on defence mechanisms of corynebacteria against bacteriophages.     

Improvement of plasmid encoded lactococcal bacteriophage resistance by mutator strain Epicurian coli XL1-Red

A random mutation strategy using mutator strain, Epicurian coli XL1-Red, was applied to a plasmid, pND018, constructed by inserting a Lactococcus lacis bacteriophage resistance gene (abiI) into a L. lactis/E. coli shuttle vector (pDL278), to introduce random mutations throughout the plasmid. Following transformation of the mutated plasmid library to a plasmid free and phage sensitive strain of L. lactis (LM0230), mutated plasmids were screened by cross-streaking and efficiency of plaquing (EOP) assays. Two strains with enhanced resistance were obtained, as well as several phage sensitive strains. Repeated transformation of the mutated plasmids to LM0230 confirmed that the observed phenotypes were caused by mutations located on the plasmids. The EOP values and plaque morphology of two enhanced phage resistance mutants were characterized at 30°C and 37°C. These results indicate that this simple procedure can be applied to generate modified plasmids with improved phage resistance, which may be of commercial value.     

CRISPR/Cas系统的分类及研究现状

CRISPR技术作为一种新型的基因编辑技术,被广泛应用于微生物、动物和植物领域。对CRISPR/Cas系统的起源、分类特点等进行了阐述,重点梳理了CRISPR/Cas系统分类的发展历程,并根据最新分类方法,介绍了不同类型的CRISPR/Cas系统的作用机制、内部不同蛋白的作用特点及应用与发展,以期探寻更多的CRISPR/Cas系统,扩大该系统的应用领域。     

Potential of AbiS as defence mechanism determined by conductivity measurement

AIM: To compare pH and conductivity used in the determination of growth in reconstituted skim milk (RSM), to determine whether the presence of one or two plasmids in Lactococcus lactis had any influence on growth, and whether AbiS improved bacteriophages resistance of L. lactis. METHODS AND RESULTS: Conductivity and pH were used to determine growth in RSM. A small increase in the generation time was found with increasing number of plasmids, while their size was unimportant. The introduction of a plasmid-encoding AbiS did only enhance the level of phage resistance significant when other plasmids encoding either AbiS1 or the restriction modification system LlaBIII was present. CONCLUSIONS: The earliest detection of growth was observed by measuring pH, rather than conductance. The plasmid-encoded AbiS system has a potential to be used as a phage resistance mechanisms in L. lactis during milk fermentations, especially when combined with other anti-phage mechanisms. SIGNIFICANCE AND IMPACT OF THE STUDY: This study widened the knowledge about the influence of plasmid introduction on the growth rate of L. lactis, which is important for the construction of new strains. The level of protection against 936 groups of phages was only significant when the mechanism was present together with the RM system LlaBIII.