Cloning of pvuI isoschizomer restriction modification system from TBP 6(7) WM clone into Escherichia coli

New, Hui San (2007) Cloning of pvuI isoschizomer restriction modification system from TBP 6(7) WM clone into Escherichia coli. [Project Paper] (Submitted)

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Abstract

Cloning possibility of Pvul isoschizomer restriction-modification system from the bacteria strain, TBP 6(7) WM into Escherichia coli were studied. The presence of Pvul isoschizomer restriction endonuclease activity in the strain studied was confirmed. Various restriction patterns were obtained from the digesting bacteria strain, TBP 6(7) WM with selected restriction enzymes. HindIII was used to cut genomic DNA of bacteria strain, TBP 6(7) WM into 2-5 kb DNA fragments, which may contain gene encoding Pvul isoschizomer restriction-modification system. The DNA fragments were recovered and inserted into linear plasmids. The recombinant plasmids were transformed into Escherichia coli, Top 10 F' to construct a plasmid library. However, the plasmid library constructed from HindIII cut DNA fragment showed no methylase activity. Thus, plasmid library construction using other restriction enzymes, including AsuNHI, BamHI, Bg!II, BseX3I, EcoRI, FauNDI, HindllI, Psp124B1, Pvull, Sall, Smal, and S/r274I may need to be explored in future screening for methylase gene. The cloning of Pvul restriction modification system will need further research using other restriction enzyme digested fragments which had been identified in this study.

Item Type:	Project Paper
Faculty:	Faculty of Agriculture and Food Sciences
Depositing User:	Mr. Azman Mohamad
Date Deposited:	17 Dec 2024 06:36
Last Modified:	17 Dec 2024 06:36
URI:	http://psaspb.upm.edu.my/id/eprint/2290

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