Human cRAF Xpress Clones
Genes of the RAF family encode three cytoplasmic serine/ threonine protein kinases that are regulated by RAS and mediate cellular responses to growth signals from the cell surface to the nuclear. CRAF, also known as Raf-1 or c-Raf, functions downstream of the Ras family of membrane associated GTPases to which it binds directly. Once activated c-RAF can activate the dual specificity protein kinases MEK1/MEK2 which in turn phosphorylate to activate the ERK1 and ERK2. Activated ERKs are pleiotropic effectors of cell physiology and play an important role in the control of gene expression involved in the cell division cycle, apoptosis, cell differentiation and cell migration.
The c-terminal kinase domain of human CRAF was cloned into the Bam HI site of pMEV-2HA vector. The insert contains the c-terminal 1032bp of CRAF that encodes amino acids residues 305-648. In K375M, the catalytically essential lysine residue 375 (counting from the initiation methionine) is mutated to a methionine. When it's transfected into mammalian cells, pMEV2HA-CRAF-K375M expresses the mutant CRAF kinase domain devoid of protein kinase activity. This mutant protein can inhibit the functions of endogenous CRAF protein (i.e. dominant negative effect).
Gene Name: Homo sapiens v-raf-1 murine leukemia viral oncogene homolog 1, (RAF1). AKA: CRAF, Raf-1, c-Raf
Specifications
| Materials Provided: | 20 ug transfection ready plasmids in 40ul TE buffer. |
| Antibiotic Selections: | Kan/Neo for kanamycin selectin in bacteria and G418 selection in mammalian cells |
| Cloning vector: | pMEV-2HA |
| 5' cloning site: | Bam HI |
| 3' cloning site: | Bam HI |
| GenBank#: | NM_002880 |
| Mutations: | K375M |
Maintenance of plasmid clones
Upon receiving of the plasmids, briefly spin the vials to collect contents to the bottom before storing at 2-8oC if will be used shortly, or -20 oC for long term storage. It is recommended that you propagate the vectors in E. coli strains that are recombinant deficient (recA) and endonuclease A-deficient (endA) (e.g. DH5α or XL1-Blue).
Human cRAF protein sequence
Human cRAF nucleotide sequence
About Xpress Clones
Xpress Clones are a collection of expression verified genes, including protein kinase genes, dominant negative mutants, consititutively active mutants and kinase deficient mutants, and other expression ready genes. Unlike other collections aiming to have more genes, we focus on the genes important to your research, the mutations with a meaning to your experiment.
If your gene of interest is not in our list, please let us know and we can clone the genes with desired mutant forms, expression verified in the organism you choose.
