Methods of genome installation in a recipient host cell

• US 9,481,880 B2
• Filed: 05/01/2008
• Issued: 11/01/2016
• Est. Priority Date: 05/01/2007
• Status: Active Grant

• Current Assignee: J. Craig Venter Institute
• Original Assignee: Synthetic Genomics, Inc.
• Inventors: Glass, John I., Alperovich, Nina, Hutchison, Clyde A. III, Lartigue, Carole, Merryman, Charles E., Vashee, Sanjay, Venter, J. Craig
• Primary Examiner(s): SINGH, ANOOP KUMAR
• Application Number:
  US12/598,414
• Publication Number:
  US 20110045592A1
• Time in Patent Office: 3,106 Days
• US Class Current: 1/1
• CPC Class Codes: C12N 15/10 Processes for the isolation, preparation or purification of DNA or RNA chemical preparation of DNA or RNA C07H21/00; preparation of non-structural polynucleotides from microorganisms or with enzymes C12P19/34
• CPC Class Codes: C12N 15/1079 Screening libraries by altering the phenotype or phenotypic trait of the host reporter assays C12N15/1086
• CPC Class Codes: C12N 15/74 Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora

Abstract

The presently disclosed invention relates to methods of installing a genome isolated from one species (the donor) into suitably prepared cells of a second species (the recipient). Introduction of the donor genetic material into the recipient host cell effectively converts the recipient host cell into a new cell that, as a result of the operation of the donated genetic material, is functionally classified as belonging to the genus and species of the donor genetic material.

Claims

1. A method for transplantation of a Mycoplasma mycoides Large Colony donor cell genome in a recipient Mycoplasma capricolum, said method comprising:

(a) isolating a closed circular double stranded donor genome from a Mycoplasma mycoides Large Colony donor cell and suspending the isolated donor genome in agarose;

(b) deproteinizing the donor genome and liberating the donor genome from the agarose;

(c) preparing a Mycoplasma capricolum as a recipient cell for introduction of the donor genome by removing or inactivating the recipient cell genome, and simultaneously introducing the isolated donor genome from step (b) into the recipient cell in presence of polyethylene glycol, wherein the donor genome phenotypically transforms the recipient cell to that of the of Mycoplasma mycoides Large Colony.

2. The method of claim 1, wherein the donor genome comprises a selection marker.

3. The method of claim 1, wherein the donor genome encodes a restriction endonuclease that recognizes sequences in the recipient genome but not the donor genome.

4. The method of claim 1, wherein the donor genome is methylated.

5. The method of claim 4, wherein the donor genome is GATC Dam methylated.