Phytase Gene Linearized Fragment Into The Corn

Phytase gene is transfered to maize shen 137 to enhance the quality of the crop, and to solve the problem of animal absorbing of nutrients and degrading of phytic acid in feeds. A transfer system without vector backbone or select gene is researched to solve the bio-safety problem of transgenic plants.A pair of primer that contains T-DNA border sequence are used to amplify the plsmid pBI121/phyA II through PCR method, and gained the liner transfer element of phytase gene flanked by T-DNA border sequence, maize is transferred with the element though pollen-tube pathway method and transformants are gained.The detect of phytase gene and report gene gus of T1 generation of trasformants shows that the foreign gene has integrated into the genome of maize. The seeds are planted and T2 progeny are gained. The detect through PCR method of phytase gene and report gene gus of T2 progeny indicates that the foreign gene is stably inherited. And the sequence of the PCR products is expectedly identical to that of its counterpart in pBI121/phyAII. The transcription of foreign gene is proved by RT-PCR. Phytase activity is detected in the transformants. GUS histochemical staining shows the expression of gus gene.