What is the DHFR gene?
DHFR (Dihydrofolate Reductase) is a Protein Coding gene. Diseases associated with DHFR include Megaloblastic Anemia Due To Dihydrofolate Reductase Deficiency and Megaloblastic Anemia. Among its related pathways are Folate Metabolism and E2F mediated regulation of DNA replication.
Does DHFR require glycosylation?
Recombinant Production of DHFR Cell Selection Also, posttranslational modifications such as phosphorylation and glycosylation are not required for the human DHFR to function properly, also making E.
What information does the DHFR enzyme activity provide in the context of recombinant protein expression and purification?
What info does the DHFR enzyme activity provide in the context of recombinant protein expression and purification? Its activity will determine how useful and effective recombinant protein purification is- and thus how useful it is.
What is the function of DHFR?
Dihydrofolate reductase (DHFR) catalyzes the reduction of dihydrofolate to tetrahydrofolate (THF). THF is needed for the action of folate-dependent enzymes and is thus essential for DNA synthesis and methylation.
What reaction does DHFR catalyze what is the cofactor that is necessary for this reaction to occur?
Does E. coli phosphorylate recombinant proteins?
Specific serine and threonine residues of recombinant human beta-casein produced in Escherichia coli were shown to be phosphorylated in vivo when human casein kinase II was coexpressed in the same plasmid. All of the phosphorylated forms found in the native protein were also detected in the recombinant protein.
How do bacteria make proteins?
Recombinant protein expression in bacteria requires the insertion of a DNA fragment ( open reading frame, ORF) into an expression vector, routinely a plasmid vector and the transferral of this vector into bacterial cells ( transformation). The cells are then cultured and induced to express the desired protein.
Can CRISPR knock out DHFR and dhfr2 in HEK293 cells?
We doubly knocked out DHFR and DHFR2 in the MAN1A1/A2/B1/C1-quadruple knockout HEK293 (QD-KO) cells, using the CRISPR/Cas9 system. The DHFR-deficient QD-KO cells were used to overexpress two proteins, lysosomal acid lipase and the constant fragment of human immunoglobulin G1by the DHFR/MTX gene-amplification method.
Can DHFR-deficient QD-KO cells be overexpressed by DHFR/MTX gene amplification?
The DHFR-deficient QD-KO cells were used to overexpress two proteins, lysosomal acid lipase and the constant fragment of human immunoglobulin G1by the DHFR/MTX gene-amplification method. This method resulted in a dramatic increase in the two protein expressions in the DHFR-deficient QD-KO cells by increasing MTX concentration.
Can human embryonic kidney 293 (HEK293) cells produce recombinant proteins?
Human embryonic kidney 293 (HEK293) cells are alternative host cells widely used for protein production. In most case, however, the cells are used for the transient expression, and there is no gene amplification system in HEK293 cells. In this study, we established a DHFR-deficient HEK293 cell line for the high yield of recombinant proteins.
Can CHO cells be used for recombinant protein production?
High productive and stable clonal cell lines for recombinant protein production have been established from the DHFR-deficient CHO cell using the dihydrofolate reductase/methotrexate (DHFR/MTX) selection methods. Human embryonic kidney 293 (HEK293) cells are alternative host cells widely used for protein production.