Abstract:
Electroporation has been used in plants and microalgae genetic engmeenng to
produce beneficial transgenic plants and microalgae. Genetic transformation by
electroporation increase the level of polyunsaturated fatty acids in Ch/ore/la
sp. such as m-3 C18:3 and m-6 C18:2. The objectives of this study are to transform Ch/ore/la
sp. with p35S-AP construct by electroporatian and to select for the putative
recombinant chlorella cells. The extracted plasmid DNA of p35S-AP was verified
using PCR technique. In this step, three types of primer combination were used as
follows: 35S-F/35S-R, PTE-VF1/PTE-VR2 and 35S-F/PTE-VF1. The 35S-F/35S-R
primer combination successfully amplified the CaMV 35S promoter with the size of
326 bp. The combination of PTE-VF1/PTE-VR2 primer successfully amplified the
antisense palmitoyl-ACP thioesterase gene fragment with the size of 617 bp while
35S-F/PTE-VF1 successfully amplified the CaMV 35S promoter and antisense
palmitoyl-ACP thioesterase gene fragment with the size of 934 bp. The DNA plasmid
was then digested with EcoRl restriction enzyme to produce linear plasmid followed
by DNA plasmid purification.
