The mariner Mos1 transposase produced in tobacco is active in vitro.
The mariner-like transposon Mos1 is used for insertional mutagenesis and transgenesis in different animals (insects, nematodes), but has never been used in plants. In this paper, the transposition activity of Mos1 was tested in Nicotiana tabacum, but no transposition event was detected. In an attempt to understand the absence of in planta transposition, Mos1 transposase (MOS1) was produced and purified from transgenic tobacco (HMNtMOS1). HMNtMOS1 was able to perform all transposition reaction steps in vitro: binding to ITR, excision and integration of the same pseudo-transposon used in in planta transposition assays. The in vitro transposition reaction was not inhibited by tobacco nuclear proteins, and did not depend on the temperature used for plant growth. Several hypotheses are proposed that could explain the inhibition of HMNtMOS1 activity in planta.
Thomas
Xavier
X
Université François Rabelais de Tours, GICC, Parc de Grandmont, 37200 Tours, France.
Hedhili
Sabah
S
Beuf
Laurent
L
Demattéi
Marie-Véronique
MV
Laparra
Hélène
H
Khong
Giang Ngan
GN
Breitler
Jean-Christophe
JC
Montandon
Frédéric
F
Carnus
Elodie
E
Norre
Frédéric
F
Burtin
Daniel
D
Gantet
Pascal
P
Bigot
Yves
Y
Renault
Sylvaine
S
D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D066298:In Vitro Techniques; D008856:Microscopy, Fluorescence / Q000379:methods; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010944:Plants / Q000235:genetics; D030821:Plants, Genetically Modified; D011485:Protein Binding; D011523:Protoplasts / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics; D013696:Temperature; D014026:Nicotiana / Q000201:enzymology* / Q000235:genetics*; D019895:Transposases / Q000235:genetics*
D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; C111652:mariner transposases; D049452:Green Fluorescent Proteins; D019895:Transposases