23. A nucleic acid vector to be used in in vivo supply of a nucleic acid of curiosity into a targeted receiver bacterial mobile, explained nucleic acid of fascination creating a specified impact on claimed specific receiver bacterial mobile, wherein claimed vector comprises: reported nucleic acid of interest, and
262、细菌噬菌体可选自长尾噬菌体科(非限制性地比如以下属：cam病毒、lika病毒、r4病毒、acadian病毒、cooper病毒、pg1病毒、pipefish病毒、rosebush病毒、brujita病毒、che9c病毒、hawkeye病毒、plot病毒、jersey病毒、k1g病毒、sp31病毒、lmd1病毒、una4病毒、bongo病毒、rey病毒、butters病毒、charlie病毒、redi病毒、baxter病毒、nymphadora病毒、bignuz病毒、fishburne病毒、phayonce病毒、kp36病毒、rogue1病毒、rtp病毒、t1病毒、tls病毒、ab18病毒、amigo病毒、anatole病毒、andromeda病毒、attis病毒、barnyard病毒、bernal13病毒、biseptima病毒、bron病毒、c2病毒、c5病毒、cba181病毒、cbast病毒、ceci病毒、che8病毒、chi病毒、cjw1病毒、corndog病毒、cronus病毒、d3112病毒、d3病毒、decurro病毒、demosthenes病毒、doucette病毒、e125病毒、eiau病毒、ff47病毒、gaia病毒、giles病毒、gordon病毒、gordtnk病毒、harrison病毒、hk578病毒、hk97病毒、jenst病毒、jwx病毒、kellezio病毒、korra病毒、l5病毒、lambda病毒、laroye病毒、liefie病毒、marvin病毒、mudcat病毒、n15病毒、nonag病毒、np1病毒、omega病毒、p12002病毒、p12024病毒、p23病毒、p70病毒、pa6病毒、pamx74病毒、tolerance病毒、pbi1病毒、pepy6病毒、pfr1病毒、phic31病毒、phicbk病毒、phieta病毒、phifel病毒、phijl1病毒、pis4a病毒、psa病毒、psimuna病毒、rdjl病毒、rer2病毒、sap6病毒、send513病毒、septima3病毒、seurat病毒、sextaec病毒、sfi11病毒、sfi21dt1病毒、sitara病毒、sk1病毒、slash病毒、smoothie病毒、soups病毒、spbeta病毒、ssp2病毒、t5病毒、tank病毒、tin2病毒、titan病毒、tm4病毒、tp21病毒、tp84病毒、tria病毒、trigintaduo病毒、vegas病毒、vendetta病毒、wbeta病毒、wildcat病毒、wizard病毒、woes病毒、xp10病毒、ydn12病毒和yua病毒)。
As is often viewed on FIGS. 10 and 11, each packaged phagemids concentrating on lacZ or stx genes are effective along with the MOIs required for killing are reminiscent of Individuals obtained with packaged phagemids containing constitutive origins of replication in the absence of antibiotic assortment. Strains not containing the target (s11983) aren't killed in any respect, as anticipated, which suggests there is very little to no nonspecific-killing.
The current inventors confirmed that it is possible to derive novel conditionally replicative plasmids, specifically based upon the primase-helicase and origin of replication from PICIs. These origins can be relatively rare in goal strains, and much more advantageously the primase-ori pair may be exceptional for each PICI, substantially cutting down the opportunity of undesired recombination or payload distribute events.
As applied herein, the conditions “phagemid” and “phasmid” are equivalent and make reference to a vector that derives from both equally a plasmid as well as a bacteriophage genome. A phagemid of your disclosure comprises a phage packaging site and an origin of replication (ori), as disclosed under.
By “donor bacterial cell” is meant herein a bacterium that is definitely effective at web hosting a vector comprising a nucleic acid of fascination, of producing a vector comprising said nucleic acid of interest and/or and that is able to transferring reported vector comprising explained nucleic acid to another bacterium. In a particular embodiment, said vector may be a phagemid, and reported donor bacterial cell could then certainly be a bacterial cell ready to make claimed phagemid, a lot more specifically in the shape of a packaged phagemid.
Preferably the genetic modification is executed inside the peptides fragment identified as epitope via the human immune process bringing about a weaker or absence of epitope recognition because of the human immune procedure.
In a specific embodiment, when explained vector is a packaged phagemid, reported vector will not comprise any component derived within the organism from which the conditional origin of replication is derived.
normally, claimed offered molecule the manufacture of which is being stopped 試玩體驗 incorporates a destructive impact on explained host organism.
In a specific embodiment, claimed natural environment could be any medium wherein explained microbiome lives, such as a stable or semi-reliable surface area or simply a liquid medium, such as drinking water, in particular squander h2o.
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In a selected embodiment, mentioned helper phage comprises a nucleic acid sequence encoding a chimeric STF comprising or consisting from the sequence SEQ ID NO: 12, reported nucleic acid sequence generally comprising or consisting of your sequence SEQ ID NO: 13, and said helper phage optionally even further comprises a nucleic acid sequence encoding a chimeric gpJ variant comprising or consisting on the sequence SEQ ID NO: 14, claimed nucleic acid sequence usually comprising or consisting on the sequence SEQ ID NO: 15.
The focused receiver bacteria can be any microorganisms, specifically existing in an organism, much more significantly in the mammal organism. It can be any commensal, symbiotic or pathogenic microorganisms of the microbiota or microbiome.