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同形小种的发现和利用:I.5+10亚基的龙麦15生物型的起源和利用 总被引:1,自引:1,他引:0
以前的SDS-PAGE分析表明龙麦15在Glu-D1位点的高分子量(HMW)麦谷蛋白亚基为2+12。我们通过对龙麦15进行大量的SDS-PAGE分析,发现了Glu-D1位点的HMW麦谷蛋白亚基为5+10的龙麦15生物型。文章分析了5+10龙麦15生物型的产生,利用及对当前优质麦育种工作的理论和实践意义。 相似文献
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小麦高分子量麦谷蛋白亚基分离方法的研究 总被引:1,自引:0,他引:1
小麦高分子量麦谷蛋白亚基(HMW-GS)与小麦面包烘烤质量和面粉的加工特性密切相关,SDS-PAGE是其常用的分离方法之一。SDS-PAGE方法一般分为2类:第一类采用11%和5%浓度的胶,后者用于分离2亚基和2^*亚基,该种方法常使用碱性提取液,需要2次电泳过程,且在5%浓度的胶中HMW-GS易于和麦醇蛋白混淆;另外一类SDS-PAGE采用梯度胶,配合使用银染方法,制梯度胶则使用梯度仪及磁力搅拌 相似文献
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“龙麦15”2+12与5+10亚基近等基因系面粉品质差异的研究 总被引:2,自引:0,他引:2
通过对小麦栽培品种龙麦15中的同形小种进行SDS-PAGE和A-PAGE的分析,在国内首次获得了一对Glu-D1位点高分子量麦谷蛋白亚基分别为2+12和5+10的近等基因系。对该近等基因系面粉品质的分析表明,带有5+10亚基的龙麦15比事有2+12亚基的龙麦15的Zeleny沉淀值高12%,沉淀值/湿面筋的比值由1.03提高到1.32。该实验结果证明,5+10亚基对面粉品质贡献确实优于2+12亚基 相似文献
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采用8-(6-氨己基)-氨基-5'-AMPSepharose亲和层析法和DEAE-Sepharose离子交换层析法从大熊猫心肌中分离纯化出了乳酸脱氢酶同工酶H4.纯化的大熊猫LDH-H4,比活为445U/mg蛋白,经SDS-PAGE,PAGE,等电聚焦电泳鉴定均为一条带,其亚基分子量为36000,等电点为5.45.经测定大熊猫LDH-H亚基N端被封闭,C端氨基酸残基经测定为Leu.氨基酸组成分析表明每个亚基含有5个Cys,9个Met. 相似文献
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人肝癌中Lewis抗原,α1,3岩藻糖转移酶与转移倾向 总被引:9,自引:0,他引:9
为了测定人原发性肝癌中Lewis抗原和α1,3岩灌糖转移酶(FucT)亚型的表达,及其与癌栓形成和转移抑制基因nm23-H1表达的关系,用免疫组化法测Lewis抗原,用Northern印迹法测FucT和NM23-H1的mRNA。结果表明,肝癌组织中SLe^x、Le^x、SDLe^x、SLe^a4和Lewis抗原表达的 率均在80%左右。其中SLe^x表达较多,Le^x和SLe^a较少,SDLe^x 相似文献
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黄淮麦区16个小麦品种(系)的Glu—1,Glu—3和Gli—1位点上的基因多样 … 总被引:7,自引:0,他引:7
16个品种(系)的高分子量麦谷蛋白亚基(HMW-GS)的等位基因频率分别为Glu-Ala(80%),Glu-Alc(12%),Glu-Alb(8%);Glu-B1b(63%)Glu-Blc(31%)Glu-Ble(6%)Glu-Dla(38%),Glu-Dld(50%)Glu-Dlc(12%),16个品种(系)的低分子量麦谷蛋白亚基(LMW-GS)的等位基因频率分别为:Glu-A3a(38%),G 相似文献
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大熊猫乳酸脱氢酶同工酶H4的分离纯化和某些性质的研究 总被引:2,自引:0,他引:2
采用8-(6-氨己基)-氨基-5-AMPSepharose亲和层分析法和DEAE-Sepharose离子交换层析法从大熊猫心肌中分离纯化出乳酸脱氢酶同工酶H4,纯化的大熊猫LDH-H4,比活为445U/mg蛋白,经SDS-PAGE,APGE,等电聚焦电泳鉴定均为一条带,其亚基分了量为36000,等电点为5.45。经测定大熊猫LDH-H亚基N端被封闭,C端氨基酸残基经测定为Leu。氨基酸组成分析表明 相似文献
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干旱对小麦幼苗诱导蛋白表达与某些生理特性的初步探讨 总被引:5,自引:0,他引:5
试验以-1.2MPaPEG6000处理动小麦种子(TriticumaestlivumL.).SDS-PAGE图谱分析表明,水分胁迫诱导幼芽及整株均产生48.4kD、41.5kD二个蛋白质亚基。在幼根中未出现以上二个蛋白亚基。胁迫48h后,根干重/芽干重比呈上升趋势,幼芽细胞膜楔对透性增大和相对含水量降幅度均大于幼根。 相似文献
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M. L. Alvarez S. Guelman N. G. Halford S. Lustig M. I. Reggiardo N. Ryabushkina P. Schewry J. Stein R. H. Vallejos 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2000,100(2):319-327
Wheat HMW glutenin subunit genes 1Ax1 and 1Dx5 were introduced, and either expressed or overexpressed, into a commercial wheat
cultivar that already expresses five subunits. Six independent transgenic events were obtained and characterized by SDS-PAGE
and Southern analysis. The 1Dx5 gene was overexpressed in two events without changes in the other endosperm proteins. Overexpression
of 1Dx5 increased the contribution of HMW glutenin subunits to total protein up to 22%. Two events express the 1Ax1 subunit
transgene with associated silencing of the 1Ax2* endogenous subunit. In the SDS-PAGE one of them shows a new HMW glutenin
band of an apparent Mr lower than that of the 1Dx5 subunit. Southern analysis of the four events confirmed transformation and suggest that the transgenes
are present in a low copy number. Silencing of all the HMW glutenin subunits was observed in two different events of transgenic
wheat expressing the 1Ax1 subunit transgene and overexpressing the Dx5 gene. Transgenes and expression patterns were stably
transmitted to the progenies in all the events except one where in some of the segregating T2 seeds the silencing of all HMW glutenin subunits was reverted associated with a drastic lost of transgenes from a high to
a low copy number. The revertant T2 seeds expressed the five endogenous subunits plus the 1Ax1 transgene.
Received: 16 June 1999 / Accepted: 29 July 1999 相似文献
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Background and Aims
The trafficking of proteins in the endoplasmic reticulum (ER) of plant cells is a topic of considerable interest since this organelle serves as an entry point for proteins destined for other organelles, as well as for the ER itself. In the current work, transgenic rice was used to study the pattern and pathway of deposition of the wheat high molecular weight (HMW) glutenin sub-unit (GS) 1Dx5 within the rice endosperm using specific antibodies to determine whether it is deposited in the same or different protein bodies from the rice storage proteins, and whether it is located in the same or separate phases within these.Methods
The protein distribution and the expression pattern of HMW sub-unit 1Dx5 in transgenic rice endosperm at different stages of development were determined using light and electron microscopy after labelling with antibodies.Key results
The use of HMW-GS-specific antibodies showed that sub-unit 1Dx5 was expressed mainly in the sub-aleurone cells of the endosperm and that it was deposited in both types of protein body present in the rice endosperm: derived from the ER and containing prolamins, and derived from the vacuole and containing glutelins. In addition, new types of protein bodies were also formed within the endosperm cells.Conclusions
The results suggest that the HMW 1Dx5 protein could be trafficked by either the ER or vacuolar pathway, possibly depending on the stage of development, and that its accumulation in the rice endosperm could compromise the structural integrity of protein bodies and their segregation into two distinct populations in the mature endosperm. 相似文献14.
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Boryana S. Stamova Ute Roessner Suganthi Suren Debbie Laudencia-Chingcuanco Antony Bacic Diane M. Beckles 《Metabolomics : Official journal of the Metabolomic Society》2009,5(2):239-252
The primary aim of this work was to evaluate potential changes in the metabolic network of transgenic wheat grain over-expressing
the high-molecular-weight (HMW) glutenin Dx5-subunit gene. GC–MS and multivariate analyses were used to compare the metabolite
profiles of developing caryopses of two independently transformed lines over-expressing Dx5 and another two independently transformed lines expressing only the selectable-marker gene (controls). Developing grain at
7, 14 and 21 Days Post-Anthesis (DPA) was studied to observe differences in metabolically active tissues. There was no distinction
between the Dx5 transformants and the controls by principal component analysis (PCA) suggesting that their metabolite compositions were similar.
Most changes in metabolite levels and starch occurred at 14 DPA but tapered off by 21 DPA. Only 3 metabolites, guanine, 4-hydroxycinnamic
acid and Unknown 071306a, were altered due to Dx5 expression after correction for false discovery rates (P < 0.0005). However, discriminant function analysis (DFA) and correlative analyses of the metabolites showed that Dx5-J, which
had the highest level of Dx5 protein in ripe caryopses, could be distinguished from the other genotypes. The second aim of
this work was to determine the influence of gene transformation on the metabolome. Cross-comparison of the transformed controls
to each other, and to the Dx5 genotypes showed that approximately 50% of the metabolic changes in the Dx5 genotypes were potentially due to variations arising from gene transformation and not from the expression of the Dx5-gene
per se. This study therefore suggests the extent to which plant transformation by biolistics can potentially influence phenotype.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
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Tiwari R Priyamvada Singh R Nainawatee HS Kumar R Tyagi BS Gupta RK Nagarajan S 《Indian journal of experimental biology》2002,40(3):309-313
Detection of 1Dx5 gene and presence of 1B/1R wheat rye translocation were studied in nineteen elite Indian wheat genotypes using AS-PCR and STS markers, respectively. Fifteen genotypes had 1B/1R translocation whereas ten showed presence of 1Dx5 gene. More than 50 per cent of the genotypes tested were found positive for both 1Dx5 and 1B/1R translocation. The results are in conformity with HMW glutenin SDS-PAGE profile for 1Dx5 and cytological observations for 1B/1R translocation. 相似文献
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Altered functional properties of tritordeum by transformation with HMW glutenin subunit genes 总被引:4,自引:0,他引:4
L. Rooke F. Barro A. S. Tatham R. Fido S. Steele F. Békés P. Gras A. Martin P. A. Lazzeri P. R. Shewry P. Barcelo 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,99(5):851-858
The high-molecular-weight (HMW) subunits of wheat glutenin are the major determinants of the gluten visco-elasticity that
allows wheat doughs to be used to make bread, pasta and other food products. In order to increase the proportions of the HMW
subunits, and hence improve breadmaking performance, particle bombardment was used to transform tritordeum, a fertile amphiploid
between wild barley and pasta wheat, with genes encoding two HMW glutenin subunits (1Ax1 and 1Dx5). Of the 13 independent
transgenic lines recovered (a transformation frequency of 1.4%) six express the novel HMW subunits at levels similar to, or
higher than, those of the endogenous subunits encoded on chromosome 1B. Small-scale mixograph analysis of T2 seeds from a line expressing the transgene for 1Dx5 indicated that the addition of novel HMW subunits can result in significant
improvements in dough strength and stability, thus demonstrating that transformation can be used to modify the functional
properties of tritordeum for improved breadmaking.
Received: 15 January 1999 / Accepted: 5 February 1999 相似文献
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Wan Y Wang D Shewry R Halford G 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2002,104(5):828-839
Analysis by SDS-PAGE of total protein fractions from single seeds of Aegilops cylindrica (genomes C and D) and Triticum timopheevi (genomes A and G) showed the presence of three bands corresponding to high molecular weight subunits of glutenin (HMW subunits) in the former and two major bands and a minor band corresponding to HMW subunits in the latter. Three Ae. cylindrica and two T. timopheevi HMW subunit gene sequences, each comprising the entire coding region, were amplified by polymerase chain reaction (PCR) and their complete nucleotide sequences determined. A combination of N-terminal amino acid sequencing of the proteins identified by SDS-PAGE and alignments of the derived amino acid sequences of the proteins encoded by the PCR products identified the Ae. cylindrica HMW subunits as 1Cx, 1Cy and 1Dy, and the T. timopheevi HMW subunits as 1Gx, 1Ax and 1Ay. It was not clear whether or not a 1Gy HMW subunit was present in T. timopheevi. The PCR products from Ae. cyclindrica were derived from 1Cy and 1Dy genes and a silent 1Dx gene containing an in-frame internal stop codon, while those from T. timopheevi were derived from 1Ax and 1Ay genes. The 1Cx, 1Gx and 1Gy sequences were not amplified successfully. The proteins encoded by the five novel genes had similar structures to previously characterized HMW subunits of bread wheat (Triticum aestivum). Differences and similarities in sequence and structure, and in the distribution of cysteine residues (relevant to the ability of HMW subunits to form high Mr polymers) distinguished the HMW subunits of x- and y-type and of each genome rather than those of the different species. There was no evidence of a change in HMW subunit expression or structure resulting from selective breeding of bread wheat. The novel 1Ax, 1Ay, 1Cy and 1Dy HMW subunits were expressed in Escherichia coli, and the expressed proteins were shown to have very similar mobilities to the endogenous HMW subunits on SDS-PAGE. The truncated 1Dx gene from Ae. cylindrica failed to express in E. coli, and no HMW subunit-related protein of the size predicted for the truncated 1Dx subunit could be identified by immunodetection in seed extracts. 相似文献
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