Title Overexpression of the Bacteriophase PRD1 DNA Polymerase
Author Guhung Jung
Address Department og Biology Education, Seoul National University
Bibliography Korean Journal of Microbiology, 30(2),141-148, 1992
Key Words PRD1, DNA polymerase, overexpression
Abstract In order to overexpress bacteriophage PRD1 DNA polymerase in E. coli cells, the 2 kb HaeII fragment was isolated from phage genomic DNA. This fragment was then cloned into pEMBL_ex3 expression vector. A specific 57bp deletion was performed by using uracil containing ss DNA and oligonucleotide spanning each region to remove an unwanted non-coding region. After this deletion, the PRD1 DNA polymerase gene is totally under the control of the vector promoter and SD sequence. Upon heat induction, a protein with an apparent size of 68 kdal was overexpressed as an active PRD1 DNA polymerase. The expression of PRD1 DNA polymerase was about 1% of total E. coli protein.
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