Transformable Rhodobacter strains, method for producing transformable Rhodobacter strains

Patent metadata

Field	Value
Publication number	US-9963709-B2
Application number	US-201213619406-A
Country	US
Kind code	B2
Filing date	Sep 14, 2012
Priority date	Sep 14, 2012
Publication date	May 8, 2018
Grant date	May 8, 2018

Abstract

The invention provides an organism for expressing foreign DNA, the organism engineered to accept standard DNA carriers. The genome of the organism codes for intracytoplasmic membranes and features an interruption in at least one of the genes coding for restriction enzymes. Further provided is a system for producing biological materials comprising: selecting a vehicle to carry DNA which codes for the biological materials; determining sites on the vehicle's DNA sequence susceptible to restriction enzyme cleavage; choosing an organism to accept the vehicle based on that organism not acting upon at least one of said vehicle's sites; engineering said vehicle to contain said DNA; thereby creating a synthetic vector; and causing the synthetic vector to enter the organism so as cause expression of said DNA.

First claim

The invention claimed is:

1. An organism for producing and sequestering heterologous biological moieties, wherein the organism is R. sphaeroides , and wherein the genome of the organism codes for intracytoplasmic membranes but is modified to not code for the restriction enzyme RshI, the organism created by the method comprising:
(a) providing R. sphaeroides wherein the genome of the R. sphaeroides codes for the intracytoplasmic membranes;
(b), modifying the genome of said R. sphaeroides a first time to not code for the restriction enzyme RshI, and
(c) modifying the genome of said R. sphaeroides a second time to produce at least one heterologous membrane protein and to sequester said at least one heterologous membrane protein within said intracytoplasmic membranes.

2. The organism of claim 1 wherein modifying the genome a first time comprises deleting the gene RSP_3759 from said R. sphaeroides ' genome.

3. The organism of claim 1 wherein the R. sphaeroides is modified the first time so as to be capable of receiving replicatable and nonreplicatable vectors to immediately produce an expression strain.

4. The organism of claim 1 wherein the genome is modified the second time using electroporation.

5. The organism of claim 1 wherein the genome is modified the second time using chemical transformation.

Assignees

• Laible Philip D
• Hanson Deborah K
• Uchicago Argonne Llc

Inventors

• Laible Philip D
• Hanson Deborah K

Classifications

• C12N9/22

  Ribonucleases {[RNase]; Deoxyribonucleases [DNase]} · CPC title

• C12P21/02

  having a known sequence of two or more amino acids, e.g. glutathione · CPC title

• C12N15/74 Primary

  Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora · CPC title