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51.
RAPD fingerprints for identification and for taxonomic studies of elite poplar (Populus spp.) clones
S. Castiglione G. Wang G. Damiani C. Bandi S. Bisoffi F. Sala 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1993,87(1-2):54-59
RAPD (Random Amplified Polymorphic DNA) fingerprints have recently been used to estimate genetic and taxonomic relationships in plants. In this study RAPD analysis was performed on 32 clones belonging to different species of the genus Populus. Of these, 25 clones are registered in several countries for commercial use and, altogether, cover almost 50% of the worlds cultivated poplars. DNA was prepared from leaves and amplified by PCR using random oligonucleotide primers. Amplification products were separated by agarose-gel electrophoresis to reveal band polymorphisms. Four primers out of the 18 tested, were selected on the basis of the number and frequency of the polymorphisms produced. With these a total of 120 different DNA bands were reproducibly obtained, 92% of which were polymorphic. The polymorphisms were scored and used in band-sharing analyses to identify genetic relationships. With a few but interesting exceptions, these are consistent with the present taxonomy of the genus Populus and with the known predigrees of cultivated poplars. Moreover, the results show that RAPD analysis allows one to discriminate among all tested clones and can, therefore, be recommended as a convenient tool to defend plant breeders rights. 相似文献
52.
Identification of RAPD markers linked to a major rust resistance gene block in common bean 总被引:6,自引:0,他引:6
S. D. Haley P. N. Miklas J. R. Stavely J. Byrum J. D. Kelly 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1993,86(4):505-512
Rust in bean (Phaseolus vulgaris L.), caused byUromyces appendiculatus (Pers.) Unger var.appendiculatus [ =U. phaseoli (Reben) Wint.], is a major disease problem and production constraint in many parts of the world. The predominant form of genetic control of the pathogen is a series of major genes which necessitate the development of efficient selection strategies. Our objective was focused on the identification of RAPD (random amplified polymorphic DNA) markers linked to a major bean rust resistance gene block enabling marker-based selection and facilitating resistance gene pyramiding into susceptible bean germplasm. Using pooled DNA samples of genotyped individuals from two segregating populations, we identified two RAPD markers linked to the gene block of interest. One such RAPD, OF10970 (generated by a 5-GGAAGCTTGG-3 decamer), was found to be closely linked (2.15±1.50 centi Morgans) in coupling with the resistance gene block. The other identified RAPD, OI19460 (generated by a 5-AATGCGGGAG-3 decamer), was shown to be more tightly linked (also in coupling) than OF10970 as no recombinants were detected among 97 BC6F2 segregating individuals in the mapping population. Analysis of a collection of resistant and susceptible cultivars and experimental lines, of both Mesoamerican and Andean origin, revealed that: (1) recombination between OF10970 and the gene block has occurred as evidenced by the presence of the DNA fragment in several susceptible genotypes, (2) recombination between OI19460 and the gene block has also occurred indicating that the marker is not located within the gene block itself, and (3) marker-facilitated selection using these RAPD markers, and another previously identified, will enable gene pyramiding in Andean germplasm and certain Mesoamerican bean races in which the resistance gene block does not traditionally exist. Observations of variable recombination among Mesoamerican bean races suggested suppression of recombination between introgressed segments and divergent recurrent backgrounds.Research supported by the Michigan Agricultural Research Station and the USDA-ARS. Mention of a trademark or a proprietary product does not constitute a guarantee or warranty of the product by the USDA and does not imply its approval to the exclusion of other products that may also be suitable 相似文献
53.
K. Wolff E. D. Schoen J. Peters-Van Rijn 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1993,86(8):1033-1037
Many conditions of the RAPD reaction procedure may influence the result. This paper presents rapid detection of influential factors with a fractional factorial experiment. A more extensive study of these factors is also presented. Polymerase brand, thermal cycler brand, annealing temperature, and primer, are important factors in obtaining good DNA yields and optimal fragment patterns. Each primer has its optimal annealing temperature, and this is not correlated with the GC content of the primer. Optimal species-primer combinations have to be found by trial and error. 相似文献
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56.
采用聚合酶链式反应(PCR)技术,对HPV-18序列中引物HP_1、HP_2之间的片段(F)进行扩增,通过两组阴、阳性对照实验证明扩增片段的特异性。用不同Mg浓度的缓冲系统进行PCR反应发现,缓冲系统中Mg浓度高低是影响HPV-18/HP_1、HP_2特异扩增的重要因素,高浓度Mg导致扩增特异性降低。对17例宫颈癌组织DNA进行PCR检测,有9例检出F片段,其检出率是53%,为HPV-18与宫颈癌的相关性提供证据。 相似文献
57.
用DNA重组法表达丙肝病毒部分NS_4-NS_5基因祁自柏,谷金莲,李河民(中国药品生物制品检定所,北京100050)丙型肝炎病毒(HCV)是一种RNA病毒,为研究其复制机理,制备检测丙肝抗体的诊断试剂,需大量纯化的各种丙肝病毒蛋白,我们在国内首次用D... 相似文献
58.
Ralph Rapley 《Molecular biotechnology》1994,2(3):295-298
The polymerase chain reaction (PCR) is a powerful core molecular biology technique, which when coupled to chain termination
sequencing allows gene and DNA sequence information to be derived rapidly. A number of modifications to the basic PCR format
have been developed in an attempt to increase amplification efficiency and the specificity of the reaction. We have applied
the use of DNA-binding protein, gene 32 protein from bacteriophage T4 (T4gp32) to increase amplification efficiency with a
number of diverse templates. In addition, we have found that using single-stranded DNA-binding protein (SSB) or recA protein
in DNA sequencing reactions dramatically increases the resolution of sequencing runs. The use of DNA-binding proteins in amplification
and sequencing may prove to be generally applicable in improving the yield and quality of a number of templates from various
sources. 相似文献
59.
Christopher M. Elvin Vicki Whan Peter W. Riddles 《Molecular & general genetics : MGG》1993,240(1):132-139
Gene fragments encoding serine proteases expressed in adult buffalo fly (Haematobia irritans exigua) were amplified from cDNA using generic oligonucleotide PCR primers, based on conserved residues surrounding the active-site His and Ser amino acids found in all serine proteases. The PCR product consisted of a broad band extending from about 450 by to 520 bp, which suggested that the PCR product actually consisted of numerous DNA fragments of slightly variable sizes. Seventeen independent clones of these fragments, each with an insert of approximately 480 bp, were digested with HaeIII. Comparison of restriction fragment patterns indicated that 13 of these clones harboured different PCR products. This was confirmed by DNA sequence analysis of 9 clones. Each of the sequenced clones contained an open reading frame which included structurally conserved regions characteristic of the serine protease superfamily. This study reveals the expression of a large and highly variable repertoire of serine proteases in adult buffalo fly. Importantly, these data also demonstrate the utility of such an approach in obtaining DNA probes for use in further investigations of gene family organization and expression, as well as providing recombinant antigens in the form of fusion proteins which may be used as candidates for vaccine production. 相似文献
60.
A panel of bovine, ovine and caprine polymorphic microsatellites 总被引:1,自引:0,他引:1
We report a set of six new bovine microsatellite polymorphisms based on (CA)n repeats. They are highly polymorphic and thus represent valuable markers for genome mapping. Four of the six are polymorphic in sheep and two are polymorphic in goats. One, which is polymorphic in cattle and sheep and apparently monomorphic in goats, is X-chromosome specific and has potential value in, for example, sex determination and detection of chimaerism. 相似文献