Maize transformation using xylose isomerase gene as a selection marker

The xylA gene, encoding xylose isomerase, was cloned as a 1342-bp BamHI/SacI fragment from the E. coli. As a selection marker, the xylA gene was fused between the enhanced CaMV 35S promoter (E35S) and terminator (35St) in pBAC413 (Fig.2). pBAC413 was constructed to prevent the expression of sbeIIb i...

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Published inZhi wu sheng li yu fen zi sheng wu xue xue bao = Journal of plant physiology and molecular biology Vol. 33; no. 6; p. 547
Main Authors Guo, Xin-Mei, Zhang, Xiao-Dong, Liang, Rong-Qi, Zhang, Li-Quan, Chen, Yao-Feng
Format Journal Article
LanguageChinese
Published China 01.12.2007
Subjects
Aldose-Ketose Isomerases - genetics
Blotting, Southern
Genome, Plant - genetics
Immunoblotting
Models, Genetic
Polymerase Chain Reaction
Transformation, Genetic - genetics
Zea mays - genetics
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Summary:The xylA gene, encoding xylose isomerase, was cloned as a 1342-bp BamHI/SacI fragment from the E. coli. As a selection marker, the xylA gene was fused between the enhanced CaMV 35S promoter (E35S) and terminator (35St) in pBAC413 (Fig.2). pBAC413 was constructed to prevent the expression of sbeIIb in maize. PDS1000/He was used to bombard maize calli, which were induced to form by the elite inbred lines. The selection was carried out on the media containing concentrations of xylose from 0 to 100%. The results showed that the media containing 50% to 100% D-xylose were better, but differed with the genotype of maize (Tables 1 and 2). Successful integration of xylA gene into the maize genome was confirmed by DNA dot blotting, PCR and PCR-Southern hybridization (Figs.4 to 6). A method was established in which transformed maize cells were successively screened on a medium containing xylose instead of antibiotic and herbicide for bio-safety.
ISSN:1671-3877