Construction of recombinant plasmids harbouring single copy genomic DNA of oil palm in Escherichia coli.

Ahmad Parveez G.K., . and Shah F.H., . Construction of recombinant plasmids harbouring single copy genomic DNA of oil palm in Escherichia coli. pp. 11-18. ISSN 0128-1828

Nuclear DNA was extracted from destarched leaves of oil palm Elaeis guineensis varieties tenera dura and pisifera and from leaves of E. oleifera. The nuclear DNA samples were digested with Pst I restriction endonuclease. It was observed that there is some C-methylation in tenera and E. oleifera but not in pisifera and dura. The presence of C-methylation in tenera was of interest and fragments generated from this digestion were utilized in the construction of the library. Restricted fragments of 0.4 to 1.0 kb were obtained by gradient ultracentrifugation in 10 percent-40 percent sucrose. These fragments were cloned into pUC19 plasmid vector and transformed into E. coli DH5 x as host. Colony hybridization and dot blot hybridization were carried out on all the clones with labelled nuclear DNA. Seventy six percent of the 250 clones analyzed contained single or low copy sequences and 24 percent contained repetitive sequences.

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Abstract

Nuclear DNA was extracted from destarched leaves of oil palm Elaeis guineensis varieties tenera dura and pisifera and from leaves of E. oleifera. The nuclear DNA samples were digested with Pst I restriction endonuclease. It was observed that there is some C-methylation in tenera and E. oleifera but not in pisifera and dura. The presence of C-methylation in tenera was of interest and fragments generated from this digestion were utilized in the construction of the library. Restricted fragments of 0.4 to 1.0 kb were obtained by gradient ultracentrifugation in 10 percent-40 percent sucrose. These fragments were cloned into pUC19 plasmid vector and transformed into E. coli DH5 x as host. Colony hybridization and dot blot hybridization were carried out on all the clones with labelled nuclear DNA. Seventy six percent of the 250 clones analyzed contained single or low copy sequences and 24 percent contained repetitive sequences.

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Item Type:	Article
Additional Information:	22 ref. Summaries (En)
AGROVOC Term:	ELAEIS GUINEENSIS
AGROVOC Term:	PALMAS OLEAGINOSAS
AGROVOC Term:	PLASMIDIOS
AGROVOC Term:	ESCHERICHIA
Depositing User:	Ms. Norfaezah Khomsan
Last Modified:	24 Apr 2025 05:56
URI:	http://webagris.upm.edu.my/id/eprint/20351

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