Fig. 5

PCR and RT-PCR analysis of FPGS Os03g02030 T-DNA insertion line 5. DNA and RNA extracted from the third leaves (2-week-old) of Dongjin wild type and FPGS Os03g02030 homozygous knockout line number 5 were analysed by PCR (a) and RT-PCR (b). aLane 1 and 2 were the result from using 3gE1_for and 3gE4_rev primers, and the product size of 991 bp was expected. The positive band on this primer pair confirmed the wild-type genotype. Lane 3 and 4 were the result of using 3gE1_for and RB_rev primers, and the expected product size was 667 bp. The positive band from this primer pair suggested the presence of the T-DNA insertion. Dongjin wild-type DNA was applied in the PCR reaction of lane 1 and 3, and FPGS Os03g02030 homozygous knockout line DNA was applied in the PCR reaction of lane 2 and 4. bLane 1 and 2 were the RT-PCR result from 3gE1_for and 3gE1 1_rev primers, and the expected product size was 1,016 bp. This primer pair was used to confirm the mRNA expression of FPGS Os03g02030 gene. Lane 3 and 4 were the RT-PCR result from the rice actin primers which the expected band size was 276 bp. Actin bands demonstrated an equal amount of loaded RNA. Dongjin wild-type DNA was applied in the RT-PCR reaction of lane 1 and 3, and FPGS Os03g02030 homozygous knockout line DNA was applied in the RT-PCR reaction of lane 2 and 4. M represents 1-kb ladder standard.