Cloning of AvaII Isoschizomers restriction-modification system from piw 6(7) wm clone into Escherichia coli

Tay, Yah Hui (2007) Cloning of AvaII Isoschizomers restriction-modification system from piw 6(7) wm clone into Escherichia coli. [Project Paper] (Submitted)

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Abstract

A study was conducted to clone the Avail isoschizomer restriction -modification system from indigenous bacteria strain, PIW 6(7) WM. Avail isoschizomer characteristic was verified by comparing the lambda DNA digestion pattern with commercial Avall restriction enzyme. Subsequently, the genomic DNA of the bacteria was extracted and digested with various commercial available restriction enzymes to obtain the digestion pattern. Restriction enzymes that can digest PIW 6(7) WM to provide DNA fragments 2 - 5 kb were identified. HindIII restriction enzyme was chosen to generate the desired DNA fragment. The fragment was successfully recovered from agarose gel, and then ligated into plasmid. The recombinant plasmids were extracted from colonies obtained and then digested with lysate of bacteria strain of PIW 6(7) WM. However, no colonies were obtained from pre-screening of methylase activity. Therefore, the cloning of restriction - modification system for PIW 6(7) WM was not successful with HindIII restriction enzyme. Others restriction enzymes which can be used for further screening of methylase activity had been identified, including AsuNHI, Bgll, EcoRI, FauNDI, and Pstl.

Item Type:	Project Paper
Faculty:	Faculty of Agriculture and Food Sciences
Depositing User:	Mr. Azman Mohamad
Date Deposited:	17 Dec 2024 07:15
Last Modified:	17 Dec 2024 07:15
URI:	http://psaspb.upm.edu.my/id/eprint/2306

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