Wheat Hmw Glutenin Subunit Dx5, + Dy10 Gene Into Maize And Its Expression

As improvement the maize quality is one of the most impantant object in maize breeding,in this study genetic engineering technology was used to introduce wheat high molecular weight gultenin subunit(HMW-GS) Dx5 and Dy10 gene into maize in order to increase its food-making quality and creat a novel germplasm for breeding.The entire zein promotor was amplified from maize through LA-PCR and sequenced.The result of DNA sequence alignment showed that the sequence of our cloned zein promotor is fully identical to the published sequence(GenBank Accession No.X63667).The wheat Dx5 gene,about 2500bp fragment,was derived from the plasmid pK+Dx5B by PCR.The sequencing results showed that the identity of our cloned DNA sequences and the published sequence(GenBank Accession No.X12928) is 99%.The wheat Dy10 gene,about 2000bp fragment,was derived from the plasmid pK+Dy10A by PCR.The sequencing results showed that the identity of our cloned DNA sequences and the published sequence(GenBank Accession No.X12929) is 99%.The xylA gene is used as selective marker gene.Two fragments of wheat entire HMW-GS gene were successfully sub-cloned into the modified vector pGREEN3 to form a stable expression construct named as pGREEN337(18.3 kb),which contains xylA gene as selection marker.Through biolistic bombardment of PDS1000/He system,expression vector pGREEN337 was transformed into maize and 59 transformed maize plantlets were regenerated.PCR and PCR-Southern results showed that the Dx5 and Dy10 gene has been integrated into genome of maize.In the future,the expression of transgene at RNA and protein level,as well as the food-making quality,will be detected.