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DNA plasmids having improved expression and stability

A stable and plasmid technology, applied in the field of DNA plasmids, can solve the problems of heterogeneity, unstable plasmid copy number, etc.

Inactive Publication Date: 2011-04-13
MERIAL LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are disadvantages regarding the use of pVR1012-based plasmids for DNA vaccination, such as plasmid instability and copy number heterogeneity

Method used

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  • DNA plasmids having improved expression and stability
  • DNA plasmids having improved expression and stability
  • DNA plasmids having improved expression and stability

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] Example 1 : Effect of Kanamycin Resistance ("KanaR") Gene Expression Efficiency on Plasmid Replication Efficiency in Bacteria

[0132] The following examples illustrate the effect of kanamycin resistance ("KanaR") gene expression efficiency on plasmid replication efficiency in bacteria.

[0133]First, the KanaR expression cassette elements, in particular one or more promoters, translation initiation and transcription termination, are analyzed. After analyzing these expression elements, these elements are modified and the effect of the modified elements on the yield of plasmid production is analyzed. Four potential promoters were identified. P3 (SEQ ID NO: 3) was identified from database and literature searches, the structure is described in GenBank AN#X00928 (SEQ ID NO: 4) but not detected by current software. P2 (SEQ ID NO: 2) was identified by bioinformatic analysis and was also found in published GenBank AN#V00359 (SEQ ID NO: 5). P1 (SEQ ID NO: 1) is an addition...

Embodiment 2

[0155] Example 2 :sequence

[0156] Definition The transposon comes from the 5' region of the Tn2350Km(r) gene of plasmid P1.

[0157] Accession X00928 (SEQ ID NO: 4)

[0158] 1 gaattcccgc agattaacgc gcataacaag cggtttactc gttttggcct gcaatgtaac

[0159] 61 ataatataca ttatgcgcac taaggtagag gcagcaagat tatgcggttt tgatcagaac

[0160] 121 tcagttaacc atcccgggat tctgctctgg cccattcagc gcagtttttt actttggatg

[0161] 181 aagttaaccc atgttatatt gcacaagata aaaatatatc atcatgaaca ataaaactgt

[0162] 241 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt

[0163] 301 gctcgaggga attc

[0164] Definition of transposon Tn903.

[0165] Accession V00359J01839 (SEQ ID NO: 5)

[0166] 1 ggctttgttg aataaatcag atttcgggta agtctccccc gtagcgggtt gtgttttcag

[0167] 61 gcaatacgca cgctttcagg catacctgct ttcgtcattt tgttcagcgc tcgtaccagg

[0168] 121 gccatagcct ccgcaacctg accatcgtag tcacgcagcg tcagtgaacc cccgaacagc

[0169] 181 tgttttaccc ggtacatcgc cgtttccgct atcgagcgac gg...

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Abstract

The present invention relates to compositions and methods to improve expression of exogenous polypeptides, such as an antigen, epitope, immunogen, peptide or polypeptide of interest. More particularly, the present invention provides for DNA plasmids with increased expression and stability in compositions and methods useful for DNA vaccines.

Description

[0001] Introduced as a reference [0002] This application is by reference to US Provisional Application 60 / 795,324, filed April 27,2006. All documents cited or referenced herein (“herein cited documents”), and all documents cited or referenced in herein cited documents, together with any manufacturer’s instructions for any product mentioned herein or in any document incorporated herein by reference , descriptions, product specifications, and product listings, are hereby incorporated by reference and may be used in the practice of the invention. field of invention [0003] In general, the present invention relates to DNA vaccines and methods of using them. More specifically, the present invention relates to DNA plasmids with improved expression and stability that can be used in DNA vaccines. Background of the invention [0004] DNA vaccines, also known as gene, plasmid or polynucleotide vaccines, represent a relatively simple and economical method for immunization against...

Claims

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Application Information

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IPC IPC(8): C12N15/63
CPCC12N15/68A61P31/12C12N15/11C12N15/63
Inventor J-C·奥堂奈特
Owner MERIAL LTD