Korean Journal of Microbiology (미생물학회지)
- Volume 30 Issue 2
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- Pages.141-148
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- 1992
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- 0440-2413(pISSN)
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- 2383-9902(eISSN)
Overexpression of the bacteriophase PRD1 DNA polymerase
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/sup ex/ 3-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.