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1.
大豆下胚轴可溶性蛋白中钙激活的蛋白激酶 总被引:6,自引:0,他引:6
大豆(Glycine m ax L.) 下胚轴可溶性蛋白提取液进行自磷酸化,以SDS-PAGE电泳分析其标记产物时发现,当有较高浓度的Ca2+ 存在于反应液中时,有一条18 kD蛋白带被高强度标记,同时也可观察到另一条标记强度不高的67 kD蛋白带. 当反应时间延长到15 或30m in 时,它们的标记强度都逐渐减弱,最终从放射自显影底片上消失;在反应液中加入钙螯合剂EGTA 时,则只有67 kD 被高强度标记;在磷酸化反应过程中加入非标记ATP,蛋白中的32P逐渐被非标记磷取代,表明反应体系处于磷酸化-脱磷酸化的平衡过程中,并有结果显示这一过程是钙依赖性的. 组蛋白H1 可以使反应进程加快,表明提取液中的蛋白激酶可以利用它作为底物. 综合结果表明,18 kD和67 kD蛋白可能是具有自磷酸化能力且对Ca2+ 敏感的蛋白激酶,它们对Ca2+ 的不同反应,使得钙信号的传递更具可控性 相似文献
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The formation of R-prime plasmids was selected in crosses involving soybean microsymbionts with genomic Tn5 insertions and carrying plasmid pJB3JI (with one IS2) copy as donors and Escherichia coli HB101 as recipient. Whereas the parent plasmid was 60 kb, recombinant plasmids between 76 kb and 121 kb were obtained. Restriction and Southern analyses confirmed the mobilization of Tn5 on four R-primes from Bradyrhizobium japonicum I-110 and on an R-prime plasmid from Rhizobium fredii HH303. The largest R-prime plasmid was obtained from the rescue of two symbiotically defective R. fredii mutant strains that required adenosine.Non-standard abbreviation TDP
transposon donor pool
Scientific article number A-4728 and contribution number 7724 of the Maryland Agricultural Experiment Station 相似文献
4.
《Process Biochemistry》2014,49(4):715-723
We evaluated the feasibility of using immobilized metal-ion affinity chromatography (IMAC) with nickel ion complexed with Tris(2-aminoethyl)amine (TREN) immobilized on agarose gel for purification of human Fab fragments by negative chromatography. Efficient purification of Fab fragments from digested human IgG (immunoglobulin G) (106.4% purity) was accomplished in Tris-HCl buffer at pH 7.5 without NaCl (based on total protein concentration and radial immunodiffusion of human Fab). A technological application of Ni(II)-TREN-agarose using non transgenic soybean protein extract spiked with human Fab fragments as feedstream was also studied. Experiments using Tris-HCl at pH 7.0 as loading buffer allowed the adsorption of almost all of the soybean proteins. Sixty-six percent of the loaded human Fab fragments were recovered in the flowthrough and washing fractions with about 90% purity. These results demonstrate that Ni(II)-TREN-agarose is a potential adsorbent for recombinant Fab fragment purification. 相似文献
5.
R. G. Palmer 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1992,85(4):389-393
Summary Conditional lethality in soybean, Glycine max (L.) Merr., occurred in F2 plants when cytoplasmicchlorophyll mutant Genetic Type T275 was the female parent and when either nuclear mutants T253 or T323 plants were the male parents. Mutant T253 [Mdh1-n (Urbana) y20 (Urbana) k2] is missing two of three mitochondrial malate dehydrogenase isozymes [Mdh1-n (Urbana)] and has yellowish-green leaves [y20 (Urbana)] and a tan-saddle pattern seed coat (k2). Mutant T323 [Mdh1-n (Ames 2) y20 (Ames 2)] also is missing two of three mitochondrial malate dehydrogenase isozymes [Mdh1-n (Ames 2)] and has yellowishgreen leaves [y20 (Ames 2)], but has yellow seed coat (K2). Mutants T275, T253, and T323 are viable both in the field and glasshouse. The genotypes cyt-Y2 Mdh1-n (Urbana) y20 (Urbana) k2/Mdh1-n (Urbana) y20 (Urbana) k2 and cyt-Y2 Mdh1-n (Ames 2) y20 (Ames 2)/Mdh1-n (Ames 2) y20 (Ames 2) are conditional lethals. These genotypes are lethal under field conditions, but plants survive in reduced light under shadecloth in the glasshouse. We do not know if their interaction with cyt-Y2 is due to Mdh1-n, y20, or Mdh1-n y20. The reciprocal cross (cyt-Y2 as male parent) gives viable genotypes. These conditional lethal genotypes should be useful for studies on the interaction between organelle and nuclear genomes.This is journal paper no. J-14777 of the Iowa Agriculture and Home Economics Experiment Station, Ames, IA 50011-1010. Project 2985 相似文献
6.
James T. Li Mark C. Swanson Roy J. Rando Patricia Wentz-Murtha Inna G. Ovsyannikova Ferran Morell Manuel Lopez Charles E. Reed 《Aerobiologia》1996,12(1):173-176
There have been reported epidemics of severe asthma in Barcelona, Spain, linked to a 10 kDa low molecular mass (LMM) allergen from soybean hulls that became airborne during unloading of ships. As a preliminary probe of the potential for dispersion of this allergen in USA cities, four automated air samplers were placed around a grain elevator in New Orleans and operated continuously from May to October 1990. The allergen was extracted from the filters and immunochemically assayed for soybean aeroallergen. On 31 separate days, the airborne allergen concentration in at least one of the samples was over 10000 U/m3 similar to those observed in Barcelona on some epidemic days. Areas North and East of the elevator were most affected. Serologie studies showed that of 50 asthmatics from New Orleans who were participants in an unrelated clinical study 4 or 8% demonstrated elevated titers of IgE antibody to LMM soybean allergen. Only 1 of 475 control sera (half of which were also asthmatic) obtained elsewhere in the US was positive for LMM soybean IgE antibody. Based on the findings in this study, there is a great possibility that on some days there is enough soybean allergen in the air and a sufficient frequency of soybean aeroallergen RAST positive asthmatics in New Orleans to warrant further investigation of the contribution of soybean aeroallergen to asthma around the port of New Orleans.Supported by NIAID # A121255. Mayo Clinic and Foundation and Minnesota Lung Association. 相似文献
7.
Yun Lian He Wei Jinshe Wang Chenfang Lei Haichao Li Jinying Li Yongkang Wu Shufeng Wang Hui Zhang Tingfeng Wang Pei Du Jianqiu Guo Weiguo Lu 《Molecular ecology resources》2019,19(6):1637-1646
Soybean cyst nematode (SCN, Heterodera glycines) is a major pest of soybean that is spreading across major soybean production regions worldwide. Increased SCN virulence has recently been observed in both the United States and China. However, no study has reported a genome assembly for H. glycines at the chromosome scale. Herein, the first chromosome‐level reference genome of X12, an unusual SCN race with high infection ability, is presented. Using whole‐genome shotgun (WGS) sequencing, Pacific Biosciences (PacBio) sequencing, Illumina paired‐end sequencing, 10X Genomics linked reads and high‐throughput chromatin conformation capture (Hi‐C) genome scaffolding techniques, a 141.01‐megabase (Mb) assembled genome was obtained with scaffold and contig N50 sizes of 16.27 Mb and 330.54 kilobases (kb), respectively. The assembly showed high integrity and quality, with over 90% of Illumina reads mapped to the genome. The assembly quality was evaluated using Core Eukaryotic Genes Mapping Approach and Benchmarking Universal Single‐Copy Orthologs. A total of 11,882 genes were predicted using de novo, homolog and RNAseq data generated from eggs, second‐stage juveniles (J2), third‐stage juveniles (J3) and fourth‐stage juveniles (J4) of X12, and 79.0% of homologous sequences were annotated in the genome. These high‐quality X12 genome data will provide valuable resources for research in a broad range of areas, including fundamental nematode biology, SCN–plant interactions and co‐evolution, and also contribute to the development of technology for overall SCN management. 相似文献
8.
Yuzhe Zhang Guanglong Li Shaowang Hu Jinfeng Liu Yushi Jiang Siyan Liu Shuyan Guan Jing Qu Dan Yao i Shi Yixuan Liu 《Phyton》2022,91(6):1183-1198
Soybean (Glycine max (L.) Merr.) is an important cultivated crop, which requires much water during its growth, and drought seriously affects soybean yields. Studies have shown that the expression of small heat shock proteins can enhance drought resistance, cold resistance and salt resistance of plants. In this experiment, soybean GmHsps_p23-like gene was successfully cloned by RT-PCR, the protein encoded by the GmHsps_p23-like gene was subjected to bioinformatics analysis, and the pCAMBIA3301-GmHsps_p23-like overexpression vector and pCBSG015-GmHsps_p23-like gene editing vector were constructed. Agrobacterium-mediated method was used to transform soybeans to obtain positive plants. RT-PCR detection, rehydration experiment and drought resistance physiological and biochemical index detection were performed on the T2 generation positive transgenic soybean plants identified by PCR and Southern hybridization. The results showed that the overexpression vector plant GmHsps_p23-like gene expression increased. After rehydration, the transgenic overexpression plants returned to normal growth, and the damage to the plants was low. After drought stress, the SOD and POD activities and the PRO content of the transgenic overexpression plants increased, while the MDA content decreased. The reverse was true for soybean plants with genetically modified editing vectors. The drought resistance of the overexpressed soybeans under drought stress was higher than that of the control group, and had a stronger drought resistance. It showed that the expression of soybean GmHsps_p23-like gene can improve the drought resistance of soybean. The cloning and functional verification of soybean GmHsps_p23-like gene had not been reported yet. This is the first time that PCR technology has been used to amplify the soybean GmHsps_p23-like gene and construct an expression vector for this gene. This research has laid the foundation for transgenic technology to improve plant drought resistance and cultivate new drought-resistant transgenic soybean varieties. 相似文献
9.
Background
Soybean cyst nematode (SCN) is the most economically devastating pathogen of soybean. Two resistance loci, Rhg1 and Rhg4 primarily contribute resistance to SCN race 3 in soybean. Peking and PI 88788 are the two major sources of SCN resistance with Peking requiring both Rhg1 and Rhg4 alleles and PI 88788 only the Rhg1 allele. Although simple sequence repeat (SSR) markers have been reported for both loci, they are linked markers and limited to be applied in breeding programs due to accuracy, throughput and cost of detection methods. The objectives of this study were to develop robust functional marker assays for high-throughput selection of SCN resistance and to differentiate the sources of resistance.Results
Based on the genomic DNA sequences of 27 soybean lines with known SCN phenotypes, we have developed Kompetitive Allele Specific PCR (KASP) assays for two Single nucleotide polymorphisms (SNPs) from Glyma08g11490 for the selection of the Rhg4 resistance allele. Moreover, the genomic DNA of Glyma18g02590 at the Rhg1 locus from 11 soybean lines and cDNA of Forrest, Essex, Williams 82 and PI 88788 were fully sequenced. Pairwise sequence alignment revealed seven SNPs/insertion/deletions (InDels), five in the 6th exon and two in the last exon. Using the same 27 soybean lines, we identified one SNP that can be used to select the Rhg1 resistance allele and another SNP that can be employed to differentiate Peking and PI 88788-type resistance. These SNP markers have been validated and a strong correlation was observed between the SNP genotypes and reactions to SCN race 3 using a panel of 153 soybean lines, as well as a bi-parental population, F5–derived recombinant inbred lines (RILs) from G00-3213 x LG04-6000.Conclusions
Three functional SNP markers (two for Rhg1 locus and one for Rhg4 locus) were identified that could provide genotype information for the selection of SCN resistance and differentiate Peking from PI 88788 source for most germplasm lines. The robust KASP SNP marker assays were developed. In most contexts, use of one or two of these markers is sufficient for high-throughput marker-assisted selection of plants that will exhibit SCN resistance.Electronic supplementary material
The online version of this article (doi:10.1186/s12864-015-1531-3) contains supplementary material, which is available to authorized users. 相似文献10.