Method of producing superoxide dismutase in a bacterial cell

Details Method of producing superoxide dismutase in a bacterial cell Method of producing superoxide dismutase in a bacterial cell

C - Chemistry, Metallurgy

12

N

195/123, 195/1.3

C12N 9/02 (2006.01)

Patent

CA 1340464

An improved vector which upon introduction into a suitable host containing the thermolabile repressor CI renders the host capable of effecting expression of a desired gene. The vector is a double-stranded DNA molecule which includes in 5' to 3' order the following: the promoter and operator P L O L from lambda bacteriophage; the N utilization site; a first restriction enzyme site which follows thereafter; a ribosomal binding site; an ATG initiation codon or DNA which is converted into an ATG initiation codon upon insertion of the desired gene into the vector; a second restriction enzme site for inserting the gene in phase with the ATG initiation codon; an origin of replication and either a gene associated with a selectable or identifiable phenotypic trait manifested when the vector is present in the host or a fragment designated cI434 which includes the gene for the cI434 repressor protein and its associated promoter and operator. The distance between the 3' end of P L O L and the 5' end of the N utilization site is less than about 80 base pairs. The distance between the 3' end of the N utilization site and the 5' end of the ribosomal binding site is less than about 300 base pairs. Vectors of the invention may also include a T1T2 rRNA transcription termination sequence located 3' of the second restriction enzyme site. The inventive vectors may have origin of replications capable of autonomous production in the host of at least 400 constitutive copies. Plasmids have been constructed from the vectors and used to produce bovine, chicken, porcine and human growth hormones, human apolipoprotein E and human superoxide dismutase and analogs thereof in host cells. Such superoxide dismutase or analogs thereof may be used to catalyze the reduction of superoxide radicals, reduce reperfusion injury, prolong the survival time of isolated organs and prevent neurological injury. Enzymatically active eucaryotic superoxide dismutase or an analog thereof may also be produced by an improved method of this invention which employs a production medium having a concentration of Cu++ greater than about about 2ppm. The invention also concerns improved methods of recovering purified enzymatically active eucaryotic superoxide dismutase or analogs thereof.

616018

Affiliated with

Also associated with

Rating

Method of producing superoxide dismutase in a bacterial cell does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Method of producing superoxide dismutase in a bacterial cell, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Method of producing superoxide dismutase in a bacterial cell will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFCA-PAI-O-1250365