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Removal of mobile genetic elements from the genome of Clostridioides difficile and the implications for the organism’s biology

Clostridioides difficile is an emerging pathogen of One Health significance. Its highly variable genome contains mobile genetic elements (MGEs) such as transposons and prophages that influence its biology. Systematic deletion of each genetic element is required to determine their precise role in C. difficile biology and contribution to the wider mobilome. Here, Tn5397 (21 kb) and ϕ027 (56 kb) were deleted from C. difficile 630 and R20291, respectively, using allele replacement facilitated by CRISPR-Cas9. The 630 Tn5397 deletant transferred PaLoc at the same frequency (1 × 10−7) as 630 harboring Tn5397, indicating that Tn5397 alone did not mediate conjugative transfer of PaLoc. The R20291 ϕ027 deletant was sensitive to ϕ027 infection, and contained two unexpected features, a 2.7 kb remnant of the mutagenesis plasmid, and a putative catalase gene adjacent to the deleted prophage was also deleted. Growth kinetics of R20291 ϕ027 deletant was similar to wild type (WT) in rich medium but marginally reduced compared with WT in minimal medium. This work indicates the commonly used pMTL8000 plasmid series works well for CRISPR-Cas9-mediated gene deletion, resulting in the largest deleted locus (56.8 kb) described in C. difficile. Removal of MGEs was achieved by targeting conjugative/integrative regions to promote excision and permanent loss. The deletants created will be useful strains for investigating Tn5397 or ϕ027 prophage contribution to host virulence, fitness, and physiology, and a platform for other mutagenesis studies aimed at functional gene analysis without native transposon or phage interference in C. difficile 630 and R20291.

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Publication Date
Sun Feb 05 2017
Journal Name
Iraqi Journal Of Market Research And Consumer Protection
STUDY THE OPTIMUM CONDITIONS FOR THE REMOVAL OF HEAVY METAL ELEMENTS FROM AQUEOUS SOLUTIONS USING CONTAMINATED BACTERIA Bacillus subtilis LOCALLY ISOLATED: STUDY THE OPTIMUM CONDITIONS FOR THE REMOVAL OF HEAVY METAL ELEMENTS FROM AQUEOUS SOLUTIONS USING CONTAMINATED BACTERIA Bacillus subtilis LOCALLY ISOLATED

We studied the effect of certain environmental conditions for removing heavy metal elements from contaminated aqueous solutions (Cd, Cu, Pb, Fe, Zn, Ni, Cr) using the bacterium Bacillus subtilis to appoint the optimal conditions for removal ,The best optimum temperature range for two isolate was 30-35○C while the hydrogen number for the maximum mineral removal range was 6-7. The best primary mineral removal was 100 mg/L, while the maximum removal for all minerals was obtained after 6 hrs of Cu element time and the maximum removal efficiency was obtained after 24 hrs of Cu element. The results have proved that the best aeration for maximum removal was obtained at rotation speed of 150 rpm/minute. Inoculums of 5ml/100ml which contained 1

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Publication Date
Thu Aug 31 2023
Journal Name
Journal Of Legal Sciences
Standard Protection for the Human Genome

The legal protection of the human genome is an ethical issue related to the future of human existence.  After the disclosure of the human genetic map, scientific practices and research related to the human genome increased, especially the negative applications of genetic engineering that violates basic human rights, especially the right to human dignity, and pose real challenges to the existence and diversity of  Present and future generations. Therefore, the existence of legal rules at the national and international levels is an absolute necessity to prohibit these threats and to arrange civil and criminal liability for those who violate them.

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Publication Date
Mon Jan 01 2018
Journal Name
Al–bahith Al–a'alami
The Interactive Relationship between the Formation Elements and their Implications in the Structure of the Television Image

The image of television dominates the cognitive and artistic motivations. It is the formulation of ideas and visions along with its documentary ability. It is the main element in television work as it is a story that is narrated in pictures. Therefore, attention to image building is a major point of gravity in the work structure as a whole. On the image is the element carrying all aesthetic and expressive values of news and information directly to the hints that work to stimulate and stir the imagination of the recipient to evoke mental images added to the visual images to deepen the meanings.
All visual arts carry elements and components that follow in a particular pattern to give special meanings and specific connotations. However,

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Publication Date
Sun Oct 01 2017
Journal Name
Journal Of Educational And Psychological Researches
The content of intermediate stage biology books for multiple intelligences

The aim of current study is to analyze the content of intermediate stage biology books based on multiple intelligences. To do this, the researcher used the descriptive analytical approach. To analyze the three books of the intermediate stage, the author adopted content analysis tool and area unit. They were exposed to group of experts in methods of teaching biology, and measurement and evaluation. The findings of the study have shown a significant difference between what was expected and the observation of the multiple intelligences for the three biology books excluded the (social, musical, and kinetic intelligence), and there is no significant difference between the analysis' result of the biology books and the expectations of b

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Publication Date
Mon Jan 02 2012
Journal Name
Journal Of The Faculty Of Medicine Baghdad
Synthetic Biology: Science of the inthinkable

During the 3rd BBIC (Biosafety and Biosecurity international conference), in Amman, 13-15 September 2011, I witnessed the most astonishing and elegant lecture by the pioneer in this field Dr. Andrew Hessel (1).

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Publication Date
Tue Aug 01 2023
Journal Name
Journal Of Ecological Engineering
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Publication Date
Sun Jul 23 2023
Journal Name
Al-rafidain Journal Of Medical Sciences ( Issn 2789-3219 )
Future of Engineered Phage Therapy for Clostridium difficile Infections

Bacteriophages have the potential to eliminate both antibiotic-resistant and sensitive bacteria; as a result, they have become a major focus of such research. In contrast to antibiotics, which assault the entire bacterial population without discrimination, bacteriophages have a limited set of characteristics that allow them to target infectious microbes while avoiding friendly species (commensal microbiota). Nevertheless, large groups of naturally occurring bacteriophages that are well-differentiated and selective for the most clinically recognized pathogenic bacterial strains are required. Utilizing genetic engineering techniques that modify the target phage genome to synthesize phages with known characteristics in a brief period o

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Publication Date
Sun Dec 01 2019
Journal Name
Journal Of Economics And Administrative Sciences
Analytical study on the implications of the GATS Agreement and its implications on the Egyptian banking sector

International trade in services is becoming increasingly important as it is an economic activity that deals with invisible trade, which has become increasingly important in the balance of international trade. The establishment of the WTO is a starting point in international trade relations. It is responsible for all aspects of international trade, , And in view of the continued increase in international trade in services, the need for more internationally recognized rules has become more urgent, especially as it has been increasingly proven that the traditional framework of public services is inadequate to operate some of the most dynamic and innovative sectors of the economy. (GATS) to be the regulatory framework for this sector

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Publication Date
Mon Sep 03 2018
Journal Name
Al-academy
The communicative implications of the uniforms

The costume is a language that has carved itself into a path that is transformed over time in line with the development of human thought. The child theater is a form of communication between the individual and his society. The costume in the child's theater is seen as a sign of personality and identity. It is a model of communication that uses many of the world systems to which it belongs. Directed to the child by invoking the creative and creative energies of the designer in the formation of the children's theater. The second topic was the communicative function of secondary signs of uniforms and patterns of their transformations. The theoretical framework of the indicators included the on the research community. And his tools in the an

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Publication Date
Wed Jun 30 2004
Journal Name
Iraqi Journal Of Chemical And Petroleum Engineering
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