TH2022A - The mutant of the non-toxic vibrio colerie - Google Patents

Abstract

The invention revealed the Vibrio Cholerae strain Ogawa 395 in a genetically stable non-toxic version. This is due to the deletion mutation of the two repeats, which is the code for A1. The peptide therefore lacks the gene sequence to decode A1 the toxic peptide. It also mentioned plasmid. And the process of preparing the said species This strain is very useful as a vaccine formulation. Can be eaten both in the form of living cells Or no life

Claims (3)

1. Vibrio cholerae ogawa 395, which is non-toxic and stable genetically derivative from v .cholerae strains with at least two copies of the ctx A gene, which encodes the code of the peptide A1, A1. Will be mutated by removing some by genetic engineering. This resulted in the loss of at least a part of the necessary ctx A gene, which contributed to the transcription of the toxic A1 peptide. 2. The Claim No. 1 strain was further demonstrated to be deficient. The initial strain of the CTx A gene of V.cholerae was codified for amino acid radicals at 10-164. 3. The claim strain 1 strain was further exhibited that this strain lacked the gene sequence. CTx A of V.cholerae Initial strain at least 80% 4. The claim strain 1 strain that was further exhibited to contain the ctx B gene that coded the 5 subunit peptide B. The first claim species was further demonstrated that the species contained the entire genome of the original V.cholerae. 6. Variants of claim 1 V.cholerae The initial strain consisted of the ctx B gene that coded the B subunit peptide and that mutation was At least, the parts of the ctx B gene required for transcription were removed to obtain the complete B subunit. 7. Bacterial strain with the gene availability, ATCC storage number 39346 or ATCC storage number 39347. 8. A stable strain formation method of vibrio cholerae Ogawa 39525 that was unable to decode the toxic peptide A1 containing The wild type of Vibrio cholerae Ogawa 395 has at least 2 duplicates of the ctx A gene on the chromosome and is able to be in the gut as well as the ability to induce immunity. A mutation by partial removal of the ctx A gene by genetic engineering resulted in the DNA sequences required for transcription, missing the toxic peptide A1, and the modified V.eholerae. In order to achieve the self-transmission of the mutation, the species partially removed it into the second iterative ctx A gene on the chromosome. 9. Method for establishing a stable strain of Vibrio cholerae Ogawa. 395 non-decoding of the toxic A1 peptide containing The first plasmid containing V.cholerae DNA, consisting of 1 ctx A gene, 1 ctx B gene, and a covered segment on both sides of the ctx A gene and of the ctx B gene. The second lasmid from the first plasmid. The second plasmid was altered by removing the ctx A gene segments necessary for the transcription of the toxic peptide A1 and the plasmid segment aggregation. The second, which includes segments The DNA encapsulates the chromosome of V.cholerae Ogawa 395 progenitor species, which contains at least two duplicates of the ctx A gene on the chromosome, and is known to belong to the gut and can induce immunity. Therefore, the result was a replacement for the ctx A gene on the chromosome of V.cholerae Ogawa 395, and the variable V.eholerae was grown. In order to achieve the self-transmission of the type mutation, partially removed into the second iteration ctx A gene 1 0. Method according to claim 9 that the second plasmid had. A marker for tracking the recombination phase 1. 1. Method according to claim 9, where the missing segment from the second plasmid consists of at least part of the ctx A gene sequence coding the amino acid radicals 10 to 164. 1 2. Method according to claim 9, where the missing segment from the second plasmid contains at least 80% of the sequence used for decoding that peptide A1 1. 3.vibrio cholerae Ogawa 395 non-toxic and stable genetic strains. The initial strain of V.cholerae Ogawa 395 comprised the ctx A gene, ctx B gene, and at least two recombinant genes in the human intestine. Where the derivative strains by genetic engineering mutate by partial removal of the two ctx A genes of the substrate. This resulted in a loss of the gene sequence required for the transcription of the toxic peptide A1 from each iteration of the ctx A gene. The resulting strains consist of the ctx B gene and the subspecies genes that enable Cling to a group of people in the intestines