Construction Of Highly Efficient Plant Expression Vector Of The Major Genes PinA And PinB For Hardness In Common Wheat

Grain hardness is an important factor in determing wheat end-use quality, and it directly affect the milling quality and procession quality. A major single locus, termed Hardness (Ha) and resided on the short arm of chromosome 5D, determines whether wheat is hard or soft textured. A breakthrough for understanding the biochemical basis for endosperm came with the discovery of a 15 kDa protein composed several components, which includes Puroindoline a and b (PINA and PINB ) directly affecting the endosperm texture. Both the lack of the pinA and mutation of the pinB are inseparably linked to hard texture grain, but their role in determination of endosperm texture needs to be clarified. Therefore, we clone the pinA and pinB genes and construct the highly efficient plant expression vector.1 ) Using the primer designed by the published sequence of the pinA and pinBgenes, clone the pinA and pinB genes and construct the clone vector pGEMPaand pGEMPb.2 ) Construct the highly expressed plant expression vector pUBPa and pUBPb,which include such induced components included in the highly effeicient plant expression vector as Ubi promoter, Ω sequence, poly(A) sequence , and the Bar cassette.