RESEARCH ARTICLE

Journal of Oil Palm Research Vol. 4 No. 1 1992 June, p. 11-18

CONSTRUCTION OF RECOMBINANT PLASMIDS HARBOURING SINGLE COPY GENOMIC DNA OF OIL PALM IN ESCHERICHIA COLI

SHAH, F H *; AHMAD PARVEEZ G K+

ABSTRACT

Nuclear DNA was eitrattedfrom 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 1.0kb were obtained by gradient ultracentrifugation in 10%-40% sucrose. These fragments were cloned into pUC19 plasmid vector and transformed into E. coli DH5 a 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% contained repetitive sequences.

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* Department of Genetics,
Faculty of Life Sciences,
University Kebangsaan Malaysia,
43600 UKM Bangi,
Selangor, Malaysia.

+ Palm Oil Research Institute of Malaysia,
P O Box 10620,
50720 Kuala Lumpur