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10.1186/s12934-020-01393-2 http://scihub22266oqcxt.onion/10.1186/s12934-020-01393-2 32620105!7332536!32620105     free     free     free Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Microb+Cell+Fact 2020 ; 19 (1): 136 Nephropedia Template TP {{tp|p=32620105|t=2020. A programmable CRISPR/Cas9-based phage defense system for Escherichia coli BL21(DE3) |pdf=|usr=}}{{32620105}} gab.com Text A programmable CRISPR/Cas9-based phage defense system for Escherichia coli BL21(DE3) JO: Microb Cell Fact http://www.kidney.de/pget.php?&tw=1&pmid=32620105 #FOAvip Escherichia coli BL21 is arguably the most popular host for industrial production of proteins, and industrial fermentations are often plagued by phage infections. The CRISPR/Cas system is guided by a gRNA to cleave a specific DNA cassette, which can be developed into a highly efficient programable phage defense system. In this work, we constructed a CRISPR/Cas system targeting multiple positions on the genome of T7 phage and found that the system increased the BL21's defense ability against phage infection. Furthermore, the targeted loci on phage genome played a critical role. For better control of expression of CRISPR/Cas9, various modes were tested, and the OD of the optimized strain BL21(pT7cas9, pT7-3gRNA, prfp) after 4 h of phage infection was significantly improved, reaching 2.0, which was similar to the control culture without phage infection. Although at later time points, the defensive ability of CRISPR/Cas9 systems were not as obvious as that at early time points. The viable cell count of the engineered strain in the presence of phage was only one order of magnitude lower than that of the strain with no infection, which further demonstrated the effectiveness of the CRISPR/Cas9 phage defense system. Finally, the engineered BL21 strain under phage attack expressed RFP protein at about 60% of the un-infected control, which was significantly higher than the parent BL21. In this work, we successfully constructed a programable CRISPR/Cas9 system to increase the ability of E. coli BL21's to defend against phage infection, and created a resistant protein expression host. This work provides a simple and feasible strategy for protecting industrial E. coli strains against phage infection. Twit Text FOAVip A programmable CRISPR/Cas9-based phage defense system for Escherichia coli BL21(DE3) ????Microb Cell Fact http://www.kidney.de/pget.php?&tw=1&pmid=32620105 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=32620105 #FOAvip English Wikipedia <ref>{{Cite pmid|32620105}}</ref> A programmable CRISPR/Cas9-based phage defense system for Escherichia coli BL21(DE3) #MMPMID32620105 Liu L; Zhao D; Ye L; Zhan T; Xiong B; Hu M; Bi C; Zhang X Microb Cell Fact 2020[Jul]; 19 (1): 136 PMID32620105show ga Escherichia coli BL21 is arguably the most popular host for industrial production of proteins, and industrial fermentations are often plagued by phage infections. The CRISPR/Cas system is guided by a gRNA to cleave a specific DNA cassette, which can be developed into a highly efficient programable phage defense system. In this work, we constructed a CRISPR/Cas system targeting multiple positions on the genome of T7 phage and found that the system increased the BL21's defense ability against phage infection. Furthermore, the targeted loci on phage genome played a critical role. For better control of expression of CRISPR/Cas9, various modes were tested, and the OD of the optimized strain BL21(pT7cas9, pT7-3gRNA, prfp) after 4 h of phage infection was significantly improved, reaching 2.0, which was similar to the control culture without phage infection. Although at later time points, the defensive ability of CRISPR/Cas9 systems were not as obvious as that at early time points. The viable cell count of the engineered strain in the presence of phage was only one order of magnitude lower than that of the strain with no infection, which further demonstrated the effectiveness of the CRISPR/Cas9 phage defense system. Finally, the engineered BL21 strain under phage attack expressed RFP protein at about 60% of the un-infected control, which was significantly higher than the parent BL21. In this work, we successfully constructed a programable CRISPR/Cas9 system to increase the ability of E. coli BL21's to defend against phage infection, and created a resistant protein expression host. This work provides a simple and feasible strategy for protecting industrial E. coli strains against phage infection. |*Bacteriophages/genetics[MESH] |*CRISPR-Cas Systems[MESH] |*Escherichia coli/genetics/virology[MESH] |Genome, Viral[MESH] |Industrial Microbiology[MESH]A programmable CRISPR/Cas9-based phage defense system for Escherichia (epmc).pdf DeepDyve Pubget Overpricing