Characterisation of the expression of a novel constitutive maize promoter in transgenic wheat and maize.
A novel constitutive promoter from the maize histone H2Bgene was recently identified. In this study, we characterised H2B promoter activity in both wheat and maize tissues using the gusA reporter gene and two synthetic versions of the pat (phosphinothricin acetyl transferase) selectable marker gene, namely mopat and popat. Analyses of transgenic plants showed that the H2B promoter is able to drive the expression of gusA to strong, constitutive levels in wheat and maize tissues. Using an H2B:mopat construct and phosphinothricin selection, we recovered transgenic wheat plants at efficiencies ranging from 0.3% to 7.4% (mean 1.6%), and the efficiency of selection ranged from 40% to 100% (mean 77.7%). In another application, H2B was combined with the maize Ubi-1 or the maize Adh-1 intron to drive the expression of mopat and popat. Transformation efficiencies with the Ubi-1 intron were between 1.4- to 16-fold greater than with the Adh-1 intron. However, the use of either of the introns was necessary for the recovery of transgenic plants. Mopat gave higher transformation efficiencies and induced higher levels of PAT protein in maize tissues than popat.
Rasco-Gaunt
S
S
DuPont Wheat Transformation Laboratory, c/o IACR-Rothamsted, Harpenden, Hertfordshire AL5 2JQ, UK. Sonriza.Rasco-Gaunt@usa.dupont.com
Liu
D
D
Li
C P
CP
Doherty
A
A
Hagemann
K
K
Riley
A
A
Thompson
T
T
Brunkan
C
C
Mitchell
M
M
Lowe
K
K
Krebbers
E
E
Lazzeri
P
P
Jayne
S
S
Rice
D
D
D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D005819:Genetic Markers / Q000235:genetics; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism; D007438:Introns / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D014170:Transformation, Genetic; D014908:Triticum / Q000235:genetics*; D003313:Zea mays / Q000235:genetics*
D005819:Genetic Markers; D006657:Histones; D000123:Acetyltransferases; C056243:phosphinothricin N-acetyltransferase; D005966:Glucuronidase