detail view molecular characterisation GMO molecular characterization details

  • GMO:

    NF872
  • Alias:

  • UID:

    OKA-NBØØ3-1
  • Tradename:

    Arctic
  • Company:

    Okanagan
  • Developer:

    Okanagan
  • Species:

    Malus x domestica
    (Apple)
  • Traits:

    • Alteration in growth, development or product quality > Reduced browning

Description:

NF872 was developed through transformation in the apple cultivar Fuji with plasmid vector GEN-03. The intended consequence of genetic modification in NF872 is to reduce the expression of the PPO gene family. Reduced gene expression should lead to lower total PPO activity and result in a nonbrowning phenotype.

The PPO suppression transgene (PGAS) consists of 394, 457, 457 and 453 bp regions of apple PPO genes (PPO2, GPO3, APO5, pSR7, respectively), placed in the sense orientation under control of the Cauliflower mosaic virus 35S promoter (P-35S CaMV) and nopaline synthase terminator (T-nos). Whole genome sequencing of NF872 precisely describes three complex insertions in chromosome 3, 13 and 17 with multiple GEN-03 fragments present in each location. There is approximately 1400 bp of vector backbone (13872–67) in chromosome 3, which does not include any functional sequences.

Links regarding to this GMO:

USDA-APHIS http://www.aphis.usda.gov/brs/aphisdocs/16_00401p.pdf

Number of integration sites: 3

Transformation technique: Agrobacterium-induced

covers integration site(s)

insert only(s)

Name: insert1_NF872
Comment: The insert1_NF872 is located in CHR3 and is comprised of three vector_GEN-03 fragments. The arrangement includes an inverted repeat structure with the PPO suppression transgenes pointing towards each other. The recombination site between the two inverted transgenes was not found amongst the sequencing reads, indicating that the recombination site is within the PPO sequences (i.e. the NOS terminators and RB sequences are missing). The insertion includes 1409 bp of vector backbone (13872 –67), which does not include any functional sequences.
Map:
First Reference Sequence:
Name: insert2_NF872
Comment: The insert2_NF872 is located in CHR13 and is comprised of two vector_GEN-03 fragments. The arrangement includes an inverted repeat structure with the PPO suppression transgenes pointing towards each other. The recombination site between the two inverted transgenes is within RB sequences. The insert2_NF872 does not include any binary vector backbone sequence.
Map:
First Reference Sequence:
Name: insert3_NF872
Comment: The insert3_NF872 is located in CHR17 and is comprised of two vector_GEN-03 fragments. The arrangement includes one partial vector fragment linked to one complete PPO suppression transgene. The insertion leaves a partial CaMV35S promoter pointing out into the apple genomic sequence. In this partial CaMV35S insertion, 127 bp of the CaMV35S core promoter are missing, including the entire proximal and medial regions and 3’ end of the distal region. The insert3_NF872 does not include any binary vector backbone sequence.
Map:
First Reference Sequence:

vector(s)

Name: vector_GEN-03
Comment: GEN-03 is based on the binary vector pBINPLUS (van Engelen et al., 1995). Vector pBINPLUS is based on the widely used binary vector BIN19 (Bevan, 1984). The complete sequence of BIN19 (U09365) is available at Genbank (Benson et al., 2005). The GEN-03 vector contains a region of DNA (T-DNA) which consists of the PPO suppression transgene and NptII selection marker flanked by Agrobacterium tumefaciens T-DNA borders. Only the DNA insert (region between left border and right border) is shown. Source: USDA-APHIS petition 16_00401p (p16-17), https://www.aphis.usda.gov/brs/aphisdocs/16_00401p.pdf
Map:
First Reference Sequence: