Mammalian expression vector pUHAB
View Patent ↗The present invention relates to the construction and utilization of a new mammalian expression vector that contains a unique multiple cloning site (MCS), designated pUHAB. The pUHAB vector comprises a high copy replication origin (ColE1), a drug resistance gene (TK-Hygromycin), and a human cytomegalovirus promoter operably associated with a unique intron (hCMV/intron). Further, pUHAB comprises a selectable marker conferring resistance to kanamycin in bacterial cells, and a phage f1(+) region. pUHAB can be used to transiently or stably express cloned genes when transfected into mammalian cells. The invention also encompasses kits and host cells and cell lines comprising pUHAB, and methods of producing a recombinant protein using pUHAB.
1. A method for producing a recombinant polypeptide in an isolated host cell, comprising introducing a polynucleotide encoding the polypeptide into a multiple cloning site of a vector, wherein the vector comprises the nucleotide sequence set forth in SEQ ID NO: 1, and introducing said vector comprising the polynucleotide encoding the polypeptide into a host cell under conditions which allow for expression of the polypeptide in said cell.
2. The method of claim 1 , further comprising purifying the polypeptide.
3. The method of claim 1 wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 5-30.
4. The method of claim 1 wherein the polypeptide is an immunoglobulin.