GMO molecular characterization details

GMO detailed information

GMO:
DHA
Alias:

B0050-027
UID:
NS-B5ØØ27-4
Tradename:
Companies/Institutions:
Nuseed Americas
Species:
Brassica napus
(canola, oilseed rape, rapeseed)
Traits:
EU authorisation:

General Information

Description:	DHA canola was produced through Agrobacterium tumefaciens mediated transformation of the cultivar AV Jade with the binary vector pJP3416_GA7-ModB . The vector was designed to convert oleic acid to DHA in canola seeds and therefore contains seven FA biosynthesis genes each incorporated in its own expression cassette including seed-specific promoters, TMV 5´UTR leader as enhancer, and seed-specific terminators : the Δ 12-desaturase gene cassette (CS-delta12D-LACKL), which is under control of the _ Linum usitatissimum_ Cnl1 promotor and terminator. the ω3-/Δ15-desaturase gene cassette (CS-omega3D-PYTIR), which is under control of the Linum usitatissimum Cnl1 promotor and terminator. the Δ6-desaturase gene cassette (CS-delta6D-MICPC), which is under control of the Linum usitatissimum Cnl2 promotor and terminator. the Δ6-elongase gene cassette (CS-delta6E-PYRCR), which is under control of the Arabidopsis thaliana FAE1 promotor and the Glycine max lectin terminator. the Δ5-elongase gene cassette ( CS-delta5E-PYRCR), which is under control of the Arabidopsis thaliana FAE1 promotor and the Glycine max Lectin terminator. theΔ5-desaturase gene cassette (CS-delta5D-REBSA), which is under control of the Brassica napus FP1 promotor and the Agrobacterium tumerfaciens NOS terminator. the Δ4-desaturase gene cassette (CS-delta4D-REBSA), which is under control of the Linum usitatissimum Cnl2 promotor and terminator. The plasmid furthermore contains the neomycin-kanamycin phosphotransferase type II (nptII) and Phosphinothricin N-acetyltransferase (PAT). DHA canola was characterized with vector targeted sequencing, whole genome sequencing (WGS) and polymerase chain reaction (PCR) amplicon sequencing. Sequencing indicated that DHA canola contained no vector backbone, no marker gene nptII or any A. tumefaciens genome sequence. DHA canola accumulates substantially higher concentrations of DHA in the seed oil compared with its recipient line AV Jade as well as other conventional canola varieties. DHA canola contains two T-DNA inserts which are both required for the desired accumulation amount of DHA. Genetic stability of DHA canola was confirmed in seven generations through single seed descent. Sources: USDA-APHIS petition 17-236-01p
Links regarding to this GMO:	USDA-APHIS http://www.aphis.usda.gov/brs/aphisdocs/17_23601p.pdf
Transformation / Modification technique:	Agrobacterium-induced

Inserts / Modification

Name:	insert_DHA (putative)
Comment:	Insert order indicates order of genetic elements as described for cassettes present in the T-DNA of plasmid pJP3416_GA7-ModB.
Map:

Genetic Elements:

Reference: USDA-APHIS

Elements:

Order	Name	Donor	Size (bp)	Position	Definition	Trait	Comment
1	V-RB-RHIRD	Agrobacterium tumefaciens	163	3398-3560
2	V-intervening-SYNTH	Synthetic	68	3561-3628
3	T-cnl2-LINUS	Linum usitatissimum	538	3629-4166	Terminator of the conlinin2 gene. Truska et a. (2003). Plant Phys and Biochem, 41(2), 141-147		anti-clockwise
4	V-intervening sequences		8	4167-4174
5	CS-delta6D-MICPC	Micromonas pusilla	1395	4175-5569	delta6 desaturase gene	Altered fatty acids and oils	anti-clockwise
6	V-intervening sequences		3	5570-5572
7	L-omega-TMV	Tobacco mosaic virus	65	5573-5637			anti-clockwise
8	V-intervening sequences		8	5638-5645
9	P-cnl2-LINUS	Linum usitatissimum	2033	5646-7678	Promoter of the conlinin2 gene. Truska et al. (2003). Plant Phys and Biochem, 41, 141-147		anti-clockwise
10	V-intervening sequences		6	7679-7684
11	P-fae1-ARATH	Arabidopsis thaliana	934	7685-8618	Promoter of the fatty acid elongase gene. Rossak et al. (2001). Plant Mol Biol, 46, 717-725		clockwise
12	L-omega-TMV	Tobacco mosaic virus	65	8619-8683			clockwise
13	V-intervening sequences		3	8684-8686
14	CS-delta5E-PYRCR	Pyramimonas cordata	807	8687-9493	delta5 elongase gene	Altered fatty acids and oils	clockwise
15	V-intervening sequences		15	9494-9508
16	T-le1-SOYBN	Glycine max	334	9509-9842	Terminator of the lectin gene. Vodkin et al (1983). Cell 34, 1023-1031		clockwise
17	P-FP1-BRANA	Brassica napus	358	9843-10200	truncated napin promoter. Stalberg et al. (1993). Plant Mol Biol, 23, 671-683		clockwise
18	L-omega-TMV	Tobacco mosaic virus	65	10201-10265			clockwise
19	V-intervening sequences		3	10266-10268
20	CS-delta5D-REBSA	Pavlova salina	1281	10269-11549	delta5 desaturase gene from Pavlova salina (also known as Rebecca salina)	Altered fatty acids and oils	clockwise
21	T-nos-RHIRD	Agrobacterium tumefaciens	255	11550-11804	3' untranslated region (3' UTR) of the nopaline synthase gene from Agrobacterium tumefaciens T-DNA. Bevan et al. (1983) Nucleic Acids Res. 11: 369-385; Fraley et al. (1983) Proc Nat L Acad Sci. 80:4803-4807		clockwise
22	E-rb7_mar-TOBAC		1168	11805-12972			clockwise
23	T-cnl1-LINUS	Linum usitatissimum	734	12973-13706	Terminator of the conlinin1 gene. Truska et a. (2003). Plant Phys and Biochem, 41(2), 141-147		anti-clockwise
24	CS-omega3D-KOMPG	Pichia pastoris	1251	13707-14957	omega3 desaturase gene (Komagataella phaffii)	Altered fatty acids and oils	anti-clockwise
25	V-intervening sequences		3	14958-14960
26	L-omega-TMV	Tobacco mosaic virus	65	14961-15025			anti-clockwise
27	P-cnl1-LINUS	Linum usitatissimum	450	15026-15475	Promoter of the conlinin1 gene. Truska et al. (2003). Plant Phys and Biochem, 41,141-147		anti-clockwise
28	P-cnl2-LINUS	Linum usitatissimum	2033	15476-17508	Promoter of the conlinin2 gene. Truska et al. (2003). Plant Phys and Biochem, 41, 141-147		clockwise
29	L-omega-TMV	Tobacco mosaic virus	65	17509-17573			clockwise
30	V-intervening sequences		3	17574-17576
31	CS-delta4D-REBSA	Pavlova salina	1347	17577-18923	delta4 desaturase gene from Pavlova salina (also known as Rebecca salina)	Altered fatty acids and oils	clockwise
32	T-cnl2-LINUS	Linum usitatissimum	538	18924-19461	Terminator of the conlinin2 gene. Truska et a. (2003). Plant Phys and Biochem, 41(2), 141-147		clockwise
33	P-cnl1-LINUS	Linum usitatissimum	450	19462-19911	Promoter of the conlinin1 gene. Truska et al. (2003). Plant Phys and Biochem, 41,141-147		clockwise
34	L-omega-TMV	Tobacco mosaic virus	65	19912-19976			clockwise
35	V-intervening sequences		3	19977-19979
36	CS-delta12D-LACKL	Lachancea kluyveri	1254	19980-21233	delta12 desaturase gene	Altered fatty acids and oils	clockwise
37	T-cnl1-LINUS	Linum usitatissimum	734	21234-21967	Terminator of the conlinin1 gene. Truska et a. (2003). Plant Phys and Biochem, 41(2), 141-147		clockwise
38	E-rb7_mar-TOBAC		1168	21968-23135			clockwise
39	V-intervening sequences		8	23136-23143
40	P-fae1-ARATH	Arabidopsis thaliana	934	23144-24077	Promoter of the fatty acid elongase gene. Rossak et al. (2001). Plant Mol Biol, 46, 717-725		clockwise
41	L-omega-TMV	Tobacco mosaic virus	65	24078-24142			clockwise
42	V-intervening sequences		3	24143-24145
43	CS-delta6E-PYRCR	Pyramimonas cordata	870	24146-25015	delta6 elongase gene	Altered fatty acids and oils	clockwise
44	T-le1-SOYBN	Glycine max	334	25016-25349	Terminator of the lectin gene. Vodkin et al (1983). Cell 34, 1023-1031		clockwise
45	V-intervening sequences		22	25350-25371
46	P-35S-CaMV	Cauliflower mosaic virus	538	25372-25909			clockwise
47	V-intervening sequences		9	25910-25918
48	CS-pat-STRVR	Streptomyces viridochromogenes	552	25919-26470	Phosphinothricin acetyltransferase gene derived from Streptomyces viridochromogenes a.k.a. pat gene; shares 85 per cent homology at the amino acid level with the bar gene. Wohlleben et al. (1988) Gene 70:25-37	Glufosinate tolerance	clockwise
49	V-intervening sequences		8	26471-26478
50	T-nos-RHIRD	Agrobacterium tumefaciens	253	26479-26731	3' untranslated region (3' UTR) of the nopaline synthase gene from Agrobacterium tumefaciens T-DNA. Bevan et al. (1983) Nucleic Acids Res. 11: 369-385; Fraley et al. (1983) Proc Nat L Acad Sci. 80:4803-4807		clockwise
51	V-intervening sequences		17	26732-26748
52	V-LB-RHIRD	Agrobacterium tumefaciens	161	26749-26909

Related DNA Sequences:

Acc.No. Reference Sequence Size (bp) Annotation

USDA-APHIS No

Acc.No.	Reference	Sequence	Size (bp)	Annotation
	USDA-APHIS	No

Detection Methods

24 results

Name	Type	Target	Description	Validation	Standardisation	Verification
QL-ELE-Patf-n2/Patr-n2	element-specific	CS-pat-STRVR	Qualitative real-time PCR (TaqMan) method for detection of the pat gene (Xu et al., 2006)	in-house validation	none	1
QL-ELE-00-001	element-specific	P-Cauliflower mosaic virus	Qualitative conventional PCR method for detection of Cauliflower mosaic virus 35S promoter (Official Collection of Methods according to § 64 LFGB, L 00.00-31)	ring trial validation	national standard	2
QL-ELE-00-004	element-specific	P-Cauliflower mosaic virus	Qualitative conventional PCR method for detection of Cauliflower mosaic virus 35S promoter (Lipp et al., 2001)	ring trial validation	ISO standard	1
QL-ELE-00-005	element-specific	P-Cauliflower mosaic virus	Qualitative conventional PCR method for detection of Cauliflower mosaic virus 35S promoter (ISO/FDIS 21569)	ring trial validation	ISO standard	2
QL-ELE-00-006	element-specific	T-nos-RHIRD	Qualitative conventional PCR method for detection of nopaline synthase terminator (Official Collection of Methods according to § 64 LFGB, L 00.00-31)	ring trial validation	national standard	2
QL-ELE-00-007	element-specific	T-nos-RHIRD	Qualitative conventional PCR method for detection of nopaline synthase terminator (EU-Project SMT4-CT96-2072:1998)	ring trial validation	national standard	2
QL-ELE-00-009	element-specific	T-nos-RHIRD	Qualitative conventional PCR method for detection of nopaline synthase terminator (Lipp et al.,2001)	ring trial validation	ISO standard	2
QL-ELE-00-011	element-specific	T-nos-RHIRD	Qualitative real-time PCR (TaqMan) method for detection of nopaline synthase terminator (ISO 21569 AMD1).	ring trial validation	ISO standard	2
QL-ELE-00-012	element-specific	E-35S-CaMV, P-Cauliflower mosaic virus, P-scp1-SYNTH	Qualitative real-time PCR (TaqMan) method for detection of Cauliflower mosaic virus 35S promoter (partim P-35S). Part of the qualitative duplex real-time PCR (TaqMan) method for detection of Cauliflower mosaic virus 35S promoter and nopaline synthase terminator (ISO 21569 AMD1)	ring trial validation	ISO standard	2
QL-ELE-00-013	element-specific	T-nos-RHIRD	Qualitative real-time PCR (TaqMan) method for detection of nopaline synthase terminator (partim T-nos). Part of the qualitative duplex real-time PCR (TaqMan) method for detection of Cauliflower mosaic virus 35S promoter and nopaline synthase terminator (partim T-nos) (ISO 21569 AMD1)	ring trial validation	ISO standard	2
QL-ELE-00-017	element-specific	P-Cauliflower mosaic virus	Qualitative real-time PCR (SYBRGreen) method for detection of P-35S CaMV (Barbau-Piednoir et al., 2014)	ring trial validation	unknown	2
QL-ELE-00-018	element-specific	T-nos-RHIRD	Qualitative real-time PCR (SYBRGreen) method for detection of T-nos-RHIRD (Barbau-Piednoir et al., 2014)	ring trial validation	unknown	1
QL-TAX-BN-003	taxon-specific	Brassica napus	Qualitative real-time PCR (SYBRGreen) method for detection of oilseed rape cruciferin A gene(Barbau-Piednoir et al., 2014)	ring trial validation	unknown	1
QL-TAX-BN-004	taxon-specific	Brassica napus	Qualitative conventional PCR method for detection of oilseed rape high mobility group protein I/Y gene (Pan et al., 2007).	ring trial validation	unknown	1
QT-ELE-NOS ter 2-5' /NOS ter 2-3'	element-specific	T-nos-RHIRD	Quantitative real-time PCR (TaqMan) method for detection of T-nos (Kodama et al., 2009)	ring trial validation	none	1
QT-ELE-P35S 1-5'/P35S 1-3'	element-specific	P-Cauliflower mosaic virus	Quantitative real-time PCR (TaqMan) method for detection of the Cauliflower mosaic virus 35S promoter (Kodama et al., 2009)	ring trial validation	national standard	1
QT-ELE-00-001	element-specific	P-Cauliflower mosaic virus	Quantitative real-time PCR (TaqMan) method for detection of Cauliflower mosaic virus 35S promoter (Feinberg et al., 2005)	ring trial validation	unknown	2
QT-ELE-00-002	element-specific	CS-pat-STRVR	Quantitative real-time PCR (TaqMan) method for detection of phosphinothricin N-acetyltransferase gene (Weighardt et al., 2004) (EURL GMFF GMOMETHODS database as of February 5, 2020)	ring trial validation	unknown	1
QT-ELE-00-004	element-specific	E-35S-CaMV, P-Cauliflower mosaic virus, P-scp1-SYNTH	Quantitative real-time PCR (TaqMan) method for detection of Cauliflower mosaic virus 35S promoter (ISO 21570 Annex B.1)	ring trial validation	ISO standard	2
QT-TAX-BN-001	taxon-specific	Brassica napus	Quantitative real-time PCR (TaqMan) method for detection of oilseed rape acyl-ACP thioesterase (FatA) gene (EU-RL GMFF GMOMETHODS database as of January 23, 2014) Estimated number of target copies per haploid genome (Jacchia et al. 2018): 1	ring trial validation	EU reference method	1
QT-TAX-BN-002	taxon-specific	Brassica napus	Quantitative real-time PCR (TaqMan) method for detection of oilseed rape cruciferin storage protein (EU-RL GMFF GMOMETHODS database as of January 27, 2014). Estimated number of target copies per haploid genome (Jacchia et al. 2018): 4	ring trial validation	EU reference method	1
QT-TAX-BN-003	taxon-specific	Brassica napus	Quantitative real-time PCR (TaqMan) method for detection of oilseed rape cruciferin A gene (Jacchia et al. 2019) (EU-RL GMFF GMOMETHODS database as of January 15, 2020)	ring trial validation	EU reference method	1
QT-TAX-BN-012	taxon-specific	Brassica napus	Quantitative real-time PCR (TaqMan) method for detection of oilseed rape cruciferin A (CruA) gene (EU-RL GMFF GMOMETHODS database as of March 06, 2014). Estimated number of target copies per haploid genome (Jacchia et al. 2018): 2	ring trial validation	EU reference method	1
QT-TAX-FatA primer1/FatA primer2	taxon-specific	Brassica napus	Quantitative real-time PCR (TaqMan) method for the detection of oilseed rape (Monsanto, 2004, A recommended procedure for real-time quantitative TaqMan PCR for Roundup Ready canola RT73). Estimated number of target copies per haploid genome (Jacchia et al. 2018): 2	in-house validation	none	1

European GMO Initiative for a Unified Database System

GMO detailed information

GMO:

Alias:

UID:

Tradename:

Companies/Institutions:

Species:

Traits:

EU authorisation:

General Information

Links regarding to this GMO:

Inserts / Modification

Transformation Vectors

Detection Methods

Reference Materials

Authorisation

Literature reference