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131.
Identification of molecular markers in soybean comparing RFLP,RAPD and AFLP DNA mapping techniques 总被引:16,自引:0,他引:16
Jhy-Jhu Lin Jonathan Kuo Jin Ma James A. Saunders Hunter S. Beard Margaret H. MacDonald William Kenworthy George N. Ude Benjamin F. Matthews 《Plant Molecular Biology Reporter》1996,14(2):156-169
Three different DNA mapping techniques—RFLP, RAPD and AFLP—were used on identical soybean germplasm to compare their ability
to identify markers in the development of a genetic linkage map. Polymorphisms present in fourteen different soybean cultivars
were demonstrated using all three techniques. AFLP, a novel PCR-based technique, was able to identify multiple polymorphic
bands in a denaturing gel using 60 of 64 primer pairs tested. AFLP relies on primers designed in part on sequences for endonuclease
restriction sites and on three selective nucleotides. The 60 diagnostic primer pairs tested for AFLP analysis each distinguished
on average six polymorphic bands. Using specific primers designed for soybean fromEco RI andMse I restriction site sequences and three selective nucleotides, as many as 12 polymorphic bands per primer could be obtained
with AFLP techniques. Only 35% of the RAPD reactions identified a polymorphic band using the same soybean cultivars, and in
those positive reactions, typically only one or two polymorphic bands per gel were found. Identification of polymorphic bands
using RFLP techniques was the most cumbersome, because Southern blotting and probe hybridization were required. Over 50% of
the soybean RFLP probes examined failed to distinguish even a single polymorphic band, and the RFLP probes that did distinguish
polymorphic bands seldom identified more than one polymorphic band. We conclude that, among the three techniques tested, AFLP
is the most useful. 相似文献
132.
Time-resolved, steady-state fluorescence and fluorescence-detected circular dichroism (FDCD) have been used to resolve the fluorescence contributions of the two tryptophan residues, Trp-13 and Trp-85, in the cyclic AMP receptor protein (CRP). The iodide and acrylamide quenching data show that in CRP one tryptophan residue, Trp-85, is buried within the protein matrix and the other, Trp-13, is moderately exposed on the surface of the protein. Fluorescence-quenching-resolved spectra show that Trp-13 has emission at about 350 nm and contributes 76–83% to the total fluorescence emission. The Trp-85, unquenchable by iodide and acrylamide, has the fluorescence emission at about 337 nm. The time-resolved fluorescence measurements show that Trp-13 has a longer fluorescence decay time. The Trp-85 exhibits a shorter fluorescence decay time. In the CRP-cAMP complex the Trp-85, previously buried in the apoprotein becomes totally exposed to the iodide and acrylamide quenchers. The FDCD spectra indicate that in the CRP-cAMP complex Trp-85 remains in the same environment as in the protein alone. It has been proposed that the binding of cAMP to CRP is accompanied by a hinge reorientation of two protein domains. This allows for penetration of the quencher molecules into the Trp-85 residue previously buried in the protein matrix. 相似文献
133.
134.
东北地区生态环境中的Se及其生态效应 总被引:3,自引:3,他引:0
对东北地区不同环境要素Se的分布与转化规律的研究表明,基岩、土壤、粮食和动物毛Se含量分别为0.12、0.150-0.540、0.0096-0.0765和0.0405-0.1414μg.g^-1;城市和农村儿童发Se含量为0.460和0.182μg.g^-1。根据不同要素Se含量和生态效应,可将该区分为Se适宜区、缺乏区和过渡区3个1级区,每个1级区又分为高Se源区的低Se源区。 相似文献
135.
136.
武汉东湖水生植被及其恢复途径探讨 总被引:4,自引:0,他引:4
1992~1993年对武汉东湖三个主要湖区(郭郑湖、汤林湖和牛巢湖)水生植被的调查表明,该湖区共有水生植物32种,优势种为大茨藻、狐尾藻、苦草和菱。金鱼藻呈不断扩大的趋势。植被类型可分为11个群丛,植被面积约为0.65km ̄2,总生物量为1236.39t(湿重),植被带状分布仅见于汤林湖北部和其他部分湖汊。汤林湖和牛巢湖水生植被正处于自然恢复演替阶段。 相似文献
137.
多异瓢虫生物学的研究 总被引:3,自引:0,他引:3
多异瓢虫在山东省德州地区一年发生4代,部分个体5代,以成虫在杂草丛内、残枝落叶及土块下越冬。在自然变温下,卵、幼虫、预蛹、蛹、成虫产卵前期和全世代的发育始点和有效积温分别为18.8±0.52℃和15.0日度、16.97±0.03℃和73.3日度、15.97±0.01℃和10.23日度、15.47±0.05℃和23.05日度、17.6±0.04℃和34.1日度、18.2±0.10℃和170.3日度。据Holing圆盘方程测定,1─4龄幼虫和产卵前期成虫日最大捕食棉蚜量依次力6.7头、36.4头、77.5头、81.3头和68.0头。 相似文献
138.
通过DNA体外重组技术,以pET-3b为表达载体,构建了重组表达质粒pET-6R(B)和PET-6R(B)4,分别编码28kD的hIL-6R配基结合区片段及其53kD的二联体蛋白,并为酶切分析和DNA序列分析所证实。SDS-PAGE分析表明,含有重组表达质粒的菌株可分别表达出28kD的蛋白rIL6R-28和53kD的rIL6R-53。重组蛋白分别占菌体总蛋白的45%和29%左右。重组蛋白主要以包涵体形式存在,Western印迹表明重组蛋白具有IL-6R的抗原性。 相似文献
139.
Enrique Galindo Guadalupe Salcedo Ma. Eugenia Ramírez 《Applied microbiology and biotechnology》1994,40(5):634-637
Xanthomonas campestris NRRL B-1459 and a variant E2, when preserved on agar slopes (transferred monthly) over 11 months did not deteriorate in their ability to produce xanthan in quantity and quality, as determined by culture in 500-ml baffled flasks. Variations between 8 and 14% (with respect to the average) in the final xanthan concentration were observed for the E2 and B-1459 strains, respectively. A wide range of final viscosities was obtained; these were consistent with the changes in gum concentration. Differences were more likely associated with differences in fermentation kinetics rather than being inherent to the strains. The rheological quality of both polysacharides was relatively constant throughout the time of culture maintenance. Preservation of these bacteria on agar slopes was an adequate method, in contrast to previous reports. In the period studied, strain E2 produced higher gum titres and slightly lower gum quality compared to strain B-1459.
Correspondence to: E. Galindo 相似文献
140.
Ma Victoria Amores Paloma Hortelano Leticia García-Salguero José A. Lupiáñez 《Molecular and cellular biochemistry》1994,137(2):117-125
We have studied the effects of the diuretics mersalyl, furosemide and ethacrynic acid on renal gluconeogenesis in isolated rat-kidney tubules and on the activities of the most important gluconeogenic and glycolytic enzymes in both fed and fasted rats. Mersalyl (15 mg.kg–1 animal weight) significantly decreased the rate of gluconeogenesis in well-fed rats (68%) as well as in 24 and 48-h fasted ones (33 and 37% respectively). This inhibition occurred when lactate, pyruvate, glycerol or fructose were used as substrates. Ethacrynic acid at a dose of 50 mg.kg–1 animal weight provoked a transient inhibition of renal glucose production by almost 20% but only in fed rats with lactate as substrate, whereas the same dose of furosemide did not affect this metabolic pathway.Parallel to these changes, mersalyl caused a significant inhibition in the maximum activity of the most important gluconeogenic enzymes, phosphoenolpyruvate carboxykinase, fructose 1,6-bisphosphatase and glucose 6-phosphatase, in both fed and fasted rats. Neither ethacrynic acid nor furosemide produced any variations in the activities of these enzymes. The activity of the glycolytic enzymes phosphofructokinase and pyruvate kinase was not modified by these diuretics. Nevertheless, the activity of the thiol-enzyme glyceraldehyde 3-phosphate dehydrogenase was severely inhibited by mersalyl and to a lesser extent by the other diuretics. This inhibition was higher in fasted than fed rats. Hence, we conclude that the inhibitory effect of mersalyl on renal gluconeogenesis is due, at least partly, to a decrease in the flux through the gluconeogenic enzymes. Blood glucose was not modified after diuretic treatment in fed animals whereas mersalyl decreased the levels of blood glucose in 24-h fasted rats. Thein vivo effects of diuretics on gluconeogenesis correlate well with the previously observedin vitro effects, although ethacrynic acid was less potent as an inhibitorin vivo, probably because of its rapid clearance.Abbreviations EDTA
ethylenediaminetetraacetic acid
- EGTA
ethyleneglycolbis (-aminoethylether) N,N,N,N-tetraacetic acid
- DTT
dithiothreitol
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid
- TRIS
2-amino-2-hydroxymethyl-1,3-propanediol
Publication No. 166 from Drogas, Tóxicos Ambientales y Metabolismo Celular Research Group, Department of Biochemistry and Molecular Biology, University of Granada, Granada, Spain 相似文献