LMW-GS Gene Clone And Sequence Analysis From Psathyrostachys Huashanica

Psathymstachys huashanica is Psathyrostachys of the Hordeae race of gramineae , is the perennial herb plant. It is one of crops Cenospecies plants, has very strong stress resistance and the liking light character. It is the wheat important wild close Cenospecies resources plants, It is not only Chinese endemic species, but also the world rare and precious species. Psathyrostachys huashanica has the function of disease-resistant, drought resisting, precocious and so on fine characteristic, so it has great significance to the new wheat's variety development, low molecular weight glutenin subunits (LMW-GS) has the important influence to the wheat quality, it may form polymer related with the gluten intensity, its allelic variation are closely linked with the bread wheat's gluten quality. In order to improve the Flour Quality of Wheat, it is practicable to uses the transgene technology. In order to provides fine genetic resources to transgene to the wheat, in this research,we clone the LMW-GS gene of Psathyrostachys huashanica by the TA cloning method ,and named this gene as HS-LMW-1,and the main result as follow:(1) HS-LMW-1 gene sequence was highest homologous to the LMW-GS gene sequence from wheat-relative species, and the deduced amino acid from HS-LMW-1 gene also was highest homologous to the LMW-GS sequence from wheat-relative species,and the peptide structure was consistent with the LMW-GS peptide structure model built by Cassidy et al.(1998),so HS-LMW-1 gene with 866 base pairs is the LMW-GS gene from Psathyrostachys huashanica.(2) LMW-GS gene has conservation not only during triticum,aiso between triticum and Psathyrostachys huashanica.(3) The LMW-GS gene from Psathyrostachys huashanica is located in Ns3 site.(4) The LMW-GS gene from Psathyrostachys huashanica has single complete open reading frame encoding a mature protein of 274, with a 17bp 5'untranslated region (5'UTR), a 24 bp 3'untranslted region(3'UTR).The protein sequence is abundant with 36.8% Lysine and 19.5% proline. The structure of protein sequence is composed of 22 amino acids of signal peptide and 13 amino acids of N-terminal do-main, ,followed with 116 amino acids of repetitive domain composed of repetitive short peptide and abundance with glutamine and proline,including 14 repeating units and 13 kinds of short peptide , in which PPFSQQQQ repeated most. At the end is 123 amino acids of C-terminal domain containing 5 cysteine residues. This peptide structure was consistent with the LMW-GS peptide structure model built by Cassidy et al.(1998).(5) The regions between 36 and 67 site, between 80 and 103 site, between 110 and 147 site were intrinsic disorder regions ; the region between 166 and 267 site was Tryp_alpha_amylregion, the regions between 6 and 21, between 7 and 20, between 217 and 227, between 218 and 225 were transmembrane domain. The secondary strucure contained 24.09% Alpha helix, 4.01% Extended strand and 71.90% random coil. The protein was located in the extra-cellular space, secretory protein .This research had important significance significant to exploit and protec Psathyrostachys huashanica, utilization of shell excellent gene resource , enrich gene germplasm resource related with triticum quality,enlarge the gene source of wheat transgene.