Cloning And Sequence Analysis Of GAPDH Promoter From Changwu 134 (Triticum Aestivum L.)

The Changwu134 wheat with drought stress were used in this paper. Genomic DNA was isolated from Changwu134 wheat seeding was treated with -1.2MPa PEG6000 solution for 48h by DNAiso Reagent Kit. Through the Genome Walking method and TA cloning, the GAPDH ( glyceraldehyde-3-phosphate dehydrogenase ) upstream fragment was obtained in the wheat seeding, and the length of the sequence is 1071bp. Meanwhile, explant of mature embryos of 4 different wheat genetypes were cultured in differention media to study the induction of the callus and genetic difference of plants regeneration from different genetypes.1. The genomic DNA was extracted meets the GAPDH promoter cloning test requirements. The length of the GAPDH promoter fragment was obtained is 1071bp. Sequence analysis indicated that this fragment had no alignment with other promoters. So we speculated that that this fragment was a new fragment.2. Analysis of target fragment in promoter prediction showed that it contained four promoter motifs and one TATA-box, seven CAAT-box, and several cis-action elements such as WRKY,MYB,MYC,GT-1,HSE and ABRE can responsive to ABA, drought, hight salt and pathology.3. The results of wheat mature embryos callus induced and differentiation indicate that 2,4-D (2 mg/L-4 mg/L) was more advantageous to the induction of callus differentiation. And callus inducement rate, quality and fresh weight haven't direct relatonship with each other.