Cloning Of Endosperm-specific Promoter From Wheat And Prokaryotic Expression Of HMW Glutenin 1Bx14 Gene

High-molecular-weight glutenins which constitute of many subunits is a kind ofimportant storage protein and occupy 10% of whole protein in seed. Plenty of studies showedthat constitution of high-molecular-weight glutenin subunit (HMW-GS)play a signification roleon Triticeae processing quality. It is an important subject that transfer gene into cultivateTriticeae to improve quality utilizing gene engineering. So receiving a quality gene andpromoter which can drive it specially expression in embryo is a precondition to qualitybreeding. Triticeae high-molecular-weight glutenins is a important seed storage protein andspecially expression in embryo. We cloned a promoter utilizing PCR technology. Sequencing,homology and alignment analysis showed that the cloned product is HMW-GS 1Dx2'spromoter, the length is 1041 base pairs and the number of Genbank is AJ577815. Linked itwith Gus gene and transferred it into Triticeae seed and slide with gene gun, only observedinstant expression product in Triticeae seed, which certificated that HMW-GS promoter candirect reporting gene especially expression in Triticeae seed. Isolation specially of Triticeae seed settle foundation to use plant as vector to produceprotein and also is precondition to improve Triticeae quality utilizing gene engineeringtechnology. During improve Triticeae quality must have some quality genes and speciallypromoters. The content and constitutes of glutenins and prolamin in Triticeae seed withprocessing quality have closed relation. During the analysis contribute of different HMW-GSto processing quality find that 1Dx5+1Dy10 is quality subunits but 1Dx2+1Dy12 is badsubunits. Some quality bread Triticeae such as xiaoyan 6 (1Bx14+1By15 and1Dx2+1Dy12)have no 1Dx5+1Dy10 but have 1Dx2+1Dy12 in our country. The analysisshowed that 1Bx14+1By15 have the same function with 1Dx5+1Dy10, so research1Bx14+1By15 have signification to quality breeding. We abundance analysis HMW-GS14gene sequence, summarize anciently experimental result and succeed received HMW-GS14gene making use of ameliorative PCR buffer system and a pair of high melting temperature(Tm)primers. Insert it into prokaryotic expression vector pET30a(+), gained recombinant plasmidpET30a-HMW and transformed pET 30a -HMW into E.coli and were expressed intentionprotein product which establish foundation to function identification HMW-GS14 and improveTriticeae utilizing gene engineering.