Electroporation of Chlorella sp. with linearized psp AP-VF2 construct

Abstract

Electroporation is an important genetic transformation tool that has been used in plant genetic engineering to generate a wide variety of fertile transgenic plants. The PSP' AP-VF2 construct carries antisense palmitoyl-ACP thioesterase cDNA driven by Sesquiterpene synthase promoter. The AP2 construct was isolated from E.coli. The purity of the DNA obtained was 1.81 and its concentration was 0.15 µg/µL. The PSP' AP-VF2 plasmid was verified by PCR technique with primer combinations of PTE-VF1/PTE-VR2 and PTE-VF1/Pro-VF2. The size of amplified bands were 617 bp and 1047 bp respectively. The desired plasmid was successfully linearized by using EcoRl restriction enzymes and verified by 1.0% agarose gel electrophoresis. Purified linearized product shows the purity of 1.91 and its concentration was 0.40 µg/µL. Different parameters were used to determine the suitable voltage for transformation using p35S-AP. Agr mode (2.2kV) shows the growth of Ch/ore/la sp.