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1.
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   

2.
四川小麦地方品种Glt—1,Gli—2和Glu—1位点的遗传多样性   总被引:3,自引:0,他引:3  
运用APAGE和SDS-PAGE方法,研究了89个四川小麦(Triticum aestivum L.)地方品种Gli-1、Gli-2、Glu-1位点的遗传多样性,在这些地方品种中,总共发现32种醇溶蛋白带型和3种高分子谷蛋白带型。在Gli-1、Gli-2和Ght-1位点上,分别检测出14.15和5个等位基因。在每一个位点上,出现频率最高的等位基因分别为Gli-Als(89%),Gli-Blh(46  相似文献   

3.
小麦HMW-GS1Dx5基因的克隆及其特异性表达   总被引:3,自引:0,他引:3  
显微切割了普通小麦钢82-122(Triticumaestivum2n=42)具有1Dx5+1Dy10亚基的1D染色体长臂端,利用PCR扩增得到了HMW-GS1Dx5亚基的5(端400bp序列片段.以此作为探针从基因的组织特异性和特定发育阶段的表达两个方面研究了HMW-GS1Dx5基因表达的规律.结果表明,干种子及萌发种子中存在此基因,而在发育的幼苗中此基因未表达.HMW-GS1Dx5基因可能从开花初期开始表达.HMW-GS1Dx5基因在籽粒成熟期表达,然而在营养器官如叶片中未表达,其表达存在组织特异性.HMW-GS1Dx5基因在蜡熟期籽粒表达水平最高,其次是乳熟期籽粒.从开花15d至蜡熟期籽粒,表达趋于增加.开花15d其mRNA水平是蜡熟期籽粒mRNA的28%,灌浆期为40%、乳熟期为72%、完熟期为54%.这为进一步研究其表达调控和改善小麦品质打下基础  相似文献   

4.
酵母菌 S H2 发酵产物( 简称 F S H2) ,用 Sephadex G75 凝胶层析和 H P L C 色谱层析分离纯化,收集到一组蛋白。该组蛋白在 P A G E 和 S D S P A G E 电泳上带型一致,无蛋白亚基,分子量范围为52 ~72 k D。凝胶上糖蛋白的特异性染色——— Schiff’s 染色显示阳性染色带;用 Lowry’s 法测蛋白和硫酸酚法测糖,显示蛋白与糖的比例约为3∶1 。该组蛋白与人α干扰素作用,增强干扰素生物学效价分别为1 .6 ~2 .8 倍和1 .4 ~4 .0 倍;经 Sephadex G75 凝胶层析的 F S H2 蛋白洗脱峰增效2 .01 ~5 .68 倍;发酵液增效1 .64 ~6 .86 倍。并证实酵母菌 S H2 增效干扰素的活性成分为52 ~72k D 分子量范围的胞外糖蛋白( 简称 Y E G Ps) 。  相似文献   

5.
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   

6.
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   

7.
日本血吸虫26kD抗原基因在BCG中的表达   总被引:5,自引:0,他引:5  
研究了外源基因日本血吸虫26kD抗原(Sj26GST)在卡介苗(bacilusCalmete-Guerin,BCG)、耻垢分枝杆菌(M.smegmatis)和大肠杆菌(E.coli)中的表达.运用重组DNA和聚合酶链反应(PCR)等分子生物学技术,以表达Sj26GST的E.colipGEX衍生质粒为模板,经PCR得到编码Sj26GST的全长cDNA片段.将其按正确的阅读框顺序,克隆到人结核杆菌热休克蛋白(heatshockprotein,HSP)70的启动子下游,再将HSP70启动子和Sj26GST基因一起亚克隆到E.coli-分枝杆菌穿梭质粒pBCG-2000中,得到E.coli-分枝杆菌穿梭表达质粒pBCG-Sj26.pBCG-Sj26电转化入BCG和M.smegmatismc2155中表达Sj26GST抗原,所表达的天然重组Sj26GST(rSj26GST)为可溶性蛋白,在SDS-PAGE上分子量为26kD处可见明显的表达蛋白带.其表达量分别占BCG和M.smegmatis菌体总蛋白的15%和10%.可见,Sj26GST基因能在BCG中高效表达.  相似文献   

8.
经SephadexG-75凝胶过滤,QAE-SephadexA-50和CM-SephadexC-25离子交换层析的步骤,从湖南产尖吻蝮(Dienagkistrodonacutus)蛇毒中纯化出两个出血毒素(DaHT-1和DaHT-2).SDS-PAGE测得分子量均为23.5kD,IEF-PAGE测得等电点分别为5.6和5.2,两者具有相似的氨基酸组成,其中酸性氨基酸(Asx,Glx)分别占23%和24%,DaHT-1和DaHT-2的最小出血剂量(MHD)分别为0.5μg和0.8μg。都具蛋白水解酶活性,无对TAME,BAEE的水解活性和PLA2酶活性.两者的蛋白水解酶活力与出血活性并非正相关.DaHT-1和DaHT-2的最适温度分别为35℃和40℃,最适pH为6-9,对热均不稳定,温度高于60℃活性完全丧失。金属离子的分析显示每摩尔毒素蛋白约含0.5mol的Zn,1mol的Ca,较多的Na、K、Mg,不含Co。  相似文献   

9.
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   

10.
卢晓风  杨星勇 《动物学报》1998,44(4):406-412
巨尾阿丽蝇幼虫肠液SDS-PAGE后,X光片显影呈现3条蛋白酶活性带。IEF后出现2条蛋白酶活性带,等电点分别为PH8.5和PH7.7。肠液经硫铵沉淀,SephadexG-75凝胶过滤,SephadexDEAEA-25离子交换和SBBI-Sepharose4B亲和层析,分离化出分子量约为14KD的巨尾阿丽蝇蛋白酶。  相似文献   

11.
对5个八倍体小冰麦种子醇溶蛋白和高分子量麦谷蛋白亚基的电泳谱带进行了分析,结果表明:八倍体小冰麦中1和中2的电泳谱带基本相同,中3、中4、中5的电泳谱带基本相同,但完全不同于中1和中2的类型。八倍体小冰麦中1和中2同天蓝冰草(Agropyronintermedium(Host)P.B.=Elytrigiaintermedia(Host)Nevski=Thinopyrumintermedium(Host)BarkwarthandDewey)在高分子量麦谷蛋白亚基上存在一条相同的谱带,在醇溶蛋白谱带上出现了小麦(TriticumaestivumL.)和冰草均没有的带型。中3、中4、中5在醇溶蛋白谱带上具有一条冰草×染色体组的特征谱带,其基因表达程度同冰草类似。从5个八倍体小冰麦种子醇溶蛋白和高分子量麦谷蛋白亚基的电泳图谱结果,分析了八倍体小冰麦染色体组构成及亲本来源,并探讨了八倍体小冰麦在优质麦育种过程中的价值。  相似文献   

12.
A barley yellow dwarf virus (BYDV)-resistant line HG295 was selected from a cross between cv. 77-5433 and Zhong 5 after extensive investigation in field, greenhouse and ELISA. Cytological analysis revealed that it was an euploid line and genetically stable. The existence of alien DNA in HG295 was identified by RAPD and Southern hybridization analyses showed that the alien DNAs came from Zhong 5 or Th. intermedium. The differences of BYDV resistance between L1 and HG295 are discussed.  相似文献   

13.
Human killer cell immunoglobulin-like receptors are expressed in natural killer cells and subsets of T lymphocytes. They regulate these cells upon interaction with human leukocyte antigen class I molecules and other ligands presented by target cells. KIR gene frequencies and haplotype distributions have been shown to differ significantly between populations from different geographical regions and ethnic origins, which relates to functional variations in the immune response. We have investigated KIR gene frequencies and genotype diversities of 15 KIR genes (KIR2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 2DS1, 2DS2, 2DS3, 2DS4, ID, 2DS5, 3DL1, 3DL2, 3DL3, 3DS1) and two pseudogenes (KIR3DP1 and 2DP1) in 120 unrelated healthy individuals of the Uygur population living in the Xinjiang autonomous region of China. All individuals were typed positive for the four framework loci KIR3DL3, 2DL4, 3DL2 and KIR3DP1, while activating genes (KIR2DS1, 2DS2, 2DS3, 2DS5 and KIR3DS1) indicated some variation in this population. KIR3DS1 was found in a higher frequency in the studied population than in other groups from China. Linkage disequilibrium among KIR genes displayed a wide range. ??2 analysis, conducted among non-ubiquitous genes, based on the KIR gene frequency data from our study population and previously published population data, revealed significant differences in the KIR2DL1, 2DL2, 2DL3, 2DL5, 3DL1, 2DS1, 2DS2, 2DS3, 2DS5, and 3DS1 genes. A neighbor-joining phylogenic tree, built using the observed carrier frequencies data of 13 KIR loci (KIR2DL1, 2DL2, 2DL3, 2DL4, 2DL5, 3DL1, 3DL2, 3DL3, 2DS1, 2DS2, 2DS3, 2DS5, and 3DS1), showed relationships between the population studied and other previously reported populations. The present study can therefore be valuable for enriching the ethnical gene information resources of the KIR gene pool, for population origin studies and for KIR-related clinical practice.  相似文献   

14.
Genomic in situ hybridization (GISH) and multicolor GISH (mcGISH) methodology were used to establish the cytogenetic constitution of five partial amphiploid lines obtained from wheat × Thinopyrum intermedium hybridizations. Line Zhong 1, 2n=52, contained 14 chromosomes from each of the wheat genomes plus ten Th. intermedium chromosomes, with one pair of A-genome chromosomes having a Th. intermedium chromosomal segment translocated to the short arm. Line Zhong 2, 2n=54, had intact ABD wheat genome chromosomes plus 12 Th. intermedium chromosomes. The multicolor GISH results, using different fluorochrome labeled Th. intermedium and the various diploid wheat genomic DNAs as probes, indicated that both Zhong 1 and Zhong 2 contained one pair of Th. intermedium chromosomes with a significant homology to the wheat D genome. High-molecular-weight (HMW) glutenin and gliadin analysis revealed that Zhong 1 and Zhong 2 had identical banding patterns that contained all of the wheat bands and a specific HMW band from Th. intermedium. Zhong 1 and Zhong 2 had good HMW subunits for wheat breeding. Zhong 3 and Zhong 5, both 2n=56, possessed no gross chromosomal aberrations or translocations that were detectable at the GISH level. Zhong 4 also had a chromosome number of 2n=56 and contained the complete wheat ABD-genome chromosomes plus 14 Th. intermedium chromosomes, with one pair of Th. intermedium chromosomes being markedly smaller. Multicolor GISH results indicated that Zhong 4 also contained two pairs of reciprocally translocated chromosomes involving the A and D genomes. Zhong 3, Zhong 4 and Zhong 5 contained a specific gliadin band from Th. intermedium. Based on the above data, it was concluded that inter-genomic transfer of chromosomal segments and/or sequence introgression had occurred in these newly synthesized partial amphiploids despite their diploid-like meiotic behavior and disomic inheritance.  相似文献   

15.
黑麦(Secale cereale L., RR)是改良普通小麦(Triticum aestivum L., AABBDD)的重要基因资源,将黑麦优异基因转移到普通小麦中,是小麦品种改良的有效途经之一。文章将四川地方品种蓬安白麦子(T. aestivum L., AABBDD) 与秦岭黑麦(S. cereale cv. Qinling, RR)杂交,染色体自动加倍获得八倍体小黑麦CD-13(AABBDDRR);通过顺序FISH和GISH分析,发现该八倍体小黑麦1RS端部与7DS的端部发生相互易位,是一个携带1RS-7DS.7DL小麦-黑麦小片段易位染色体的八倍体小黑麦。利用八倍体小黑麦CD-13与四川推广小麦品种川麦42杂交、连续自交,获得包含60个株系的F5群体;对F5群体的58个株系进行GISH和FISH分析发现,其中13个株系含有1RS-7DS.7DL小片段易位染色体。在这13个株系中,株系811染色体数目为2n=6x=42,是稳定的1RS-7DS.7DL小片段易位系;并且1RS特异分子标记和醇溶蛋白分析表明,1RS-7DS.7DL易位染色体1RS小片段的断裂点位于分子标记IB267-IAG95之间,不包含编码黑麦碱蛋白的Sec-1位点;同时1RS-7DS.7DL小片段易位系的千粒重与川麦42相当,远远高于八倍体小黑麦CD-13,对千粒重无负作用。因此,1RS-7DS.7DL小麦-黑麦小片段易位系可作为进一步深入研究1RS小片段上的优异基因及其遗传效应的重要材料。  相似文献   

16.
Genomic in situhybridization (GISH) to root-tip cells at mitotic metaphase, using genomic DNA probes from Thinopyrum intermedium and Pseudoroegneria strigosa, was used to examine the genomic constitution of Th. intermedium, the 56-chromosome partial amphiploid to wheat called Zhong 5 and disease-resistant derivatives of Zhong 5, in a wheat background. Evidence from GISH indicated that Th. intermedium contained seven pairs of St, seven JS and 21 J chromosomes; three pairs of Th. intermedium chromosomes with satellites in their short arms belonging to the St, J, J genomes and homoeologous groups 1, 1, and 5 respectively. GISH results using different materials and different probes showed that seven pairs of added Th. intermedium chromosomes in Zhong 5 included three pairs of St chromosomes, two pairs of JS chromosomes and two pairs of St-JS reciprocal tanslocation chromosomes. A pair of chromosomes, which substituted a pair of wheat chromosomes in Yi 4212 and in HG 295 and was added to 21 pairs of wheat chromosomes in the disomic additions Z1, Z2 and Z6, conferred BYDV-resistance and was identical to a pair of St-JS tanslocation chromosomes (StJS) in Zhong 5. The StJS chromosome had a special GISH signal pattern and could be easily distinguished from other added chromosomes in Zhong 5; it has not yet been possible to locate the BYDV-resistant gene(s) of this translocated chromosome either in the St chromosome portion belonging to homoeologous group 2 or in the JS chromosome portion whose homoeologous group relationship is still uncertain. Among 22 chromosome pairs in disomic addition line Z3, the added chromosome pair had satellites and belonged to the St genome and homoeologous group 1. Disomic addition line Z4 carried a pair of added chromosomes which was composed of a group-7 JS chromosome translocated with a wheat chromosome; this chromosome was different to 7 Ai-1, but was identical to 7 Ai-2. The leaf rust and stem rust resistance genes were located in the distal region of the long arm, whereas the stripe rust resistance gene(s) was located in the short arm or in the proximal region of the long arm of 7 Ai-2. A pair of JS-wheat translocation chromosomes, which originated from the WJS chromosomes in Z4, was added to the disomic addition line Z5; the added chromosomes of Z5 carried leaf and stem rust resistance but not stripe rust resistance; Z5 is a potentially useful source for rust resistance genes in wheat breeding and for cloning these novel rust-resistant genes. GISH analysis using the St genome as a probe has proved advantageous in identifying alien Th. intermedium in wheat. Received: 17 May 1999 / Accepted: 22 June 1999  相似文献   

17.
小偃麦附加系Z1和Z2中外源染色体2Ai-2的结构组成@张增燕$中国农业科学院作物育种栽培研究所!北京100081@辛志勇$中国农业科学院作物育种栽培研究所!北京100081@陈孝$中国农业科学院作物育种栽培研究所!北京100081小偃麦;;附加系;;染色体  相似文献   

18.
The high-molecular-weight (HMW) glutenin genes, located on the group 1L chromosome arms, are a major determinant for baking quality in wheat ( Triticum aestivum L.). In addition, the HMW glutenin genes provide a valuable model system for studying the evolution and regulation of orthologous and paralogous genes in polyploid species. The goal of this study was to identify loci that modify the expression of the HMW glutenins, and to map them to specific chromosome arms. Comparisons were made between endosperms with zero versus three (or three versus six) doses for each of the 42 chromosome arms of wheat. SDS-PAGE and scanning densitometry were used to quantify the protein expression levels of the four HMW glutenin genes in cv. Chinese Spring, for each of the dosage comparisons. Fifteen chromosome arms were found to have significant effects on Glu-B1-1, excluding the structural gene dosage effect: eight positive effects on 1AL, 2AS, 2BL, 2DS, 5DS, 6AL, 6DL, and 7AL and seven negative effects on 1BS, 1DS, 1DL, 4DL, 6BS, 6DS, and 7AS. Nineteen chromosome arms had significant effects on Glu-B1-2, excluding the structural gene dosage effect: eight positive effects on 1AL, 2AS, 2BS, 3AL, 4BL, 6DS, 7BL and 7DS and 11 negative effects on 1AS, 1BS, 1DS, 1DL, 2AL, 2BL, 3DS, 4BS, 4DL, 5BL, and 6BS. Twenty chromosome arms had significant effects on Glu-D1-1, excluding the structural gene dosage effect: 11 positive effects on 1AL, 1BL, 2BS, 2DS, 5BS, 5DS, 6AL, 6DS, 6DL, 7AL, and 7BL and nine negative effects on 1AS, 1BS, 1DS, 2BL, 4DL, 5BL, 5DL, 6BL, and 7DS. Twenty-five chromosome arms had significant effects on Glu-D1-2, excluding the structural gene dosage effect: 17 positive effects on 1BL, 2AS, 2BS, 2DS, 2DL, 3AS, 3AL, 3BS, 5AS, 5BS, 5DL, 6AL, 6DL, 7AL, 7BS, 7BL, and 7DL and eight negative effects on 1DS, 4DL, 5AL, 5BL, 6BS, 6BL, 6DS and 7DS. Of the 164 gene-chromosome arm tests performed, about 52% (85/164) showed no significant effects, and 48% (79/164) showed significant effects, excluding the structural gene dosage effects. Of the significant effects, 56% (44/79) were positive effects, and 44% (35/79) were negative effects. Comparisons of dosage effects on orthologous loci (both x-type or both y-type HMW glutenins) showed that orthologous HMW glutenin genes are largely influenced by the same regulatory systems. Less correlation was found for comparisons between paralogous genes, although considerable conservation was observed at this level as well. These observations suggest that after polyploidization, many of the duplicated orthologous regulatory loci were inactivated by mutation, thus consolidating control over the HMW glutenin genes. Possible candidates for orthologous regulatory genes were identified in maize and barley. This study represents the first comprehensive search of the wheat genome for regulators of the HMW glutenins.  相似文献   

19.
20.
KIR2DL5 alleles were physically linked to alleles at adjacent KIR loci to define this region of KIR haplotypes in 55 gene-positive random African Americans. The majority carried KIR2DL5B. Three KIR2DL5A and six KIR2DL5B alleles that have been previously described and 11 novel KIR2DL5 alleles were identified by DNA sequencing. Novel alleles included variation that may impact promoter activity; two alleles carried nonsynonymous coding region variation. Based on linkage with KIR2DS1, KIR2DS3, KIR2DS5, KIR2DL2, KIR2DL3, and KIR3DS1 alleles, seven haplotypes of KIR2DL5A and 23 haplotypes of KIR2DL5B were observed. The phylogenetic relationships among the KIR2DL5 alleles predicted their association with either KIR2DS3 (six alleles) or KIR2DS5 (seven alleles). All of the KIR2DL5A alleles were linked either to KIR3DS1*01301 or KIR3DS1*049N. The majority of the KIR2DL5B alleles were linked to seven KIR2DL2 alleles; two were linked to a novel allele of KIR2DL3. These findings underscore the diversity of KIR haplotypes present in this population.  相似文献   

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