共查询到20条相似文献,搜索用时 15 毫秒
1.
2.
Domon E Fuijita M Ishikawa N 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2002,104(1):132-138
The length polymorphism in the waxy gene, which encodes a granule-bound ADP-glucose-glucosyl transferase [granule-bound starch
synthase I (GBSS I), E.C. 2.4.1.11] in barley (Hordeum vulgare), was found. The 5′ leader sequence of the waxy gene of barley germplasm from Japan and Korea was analyzed by the polymerase
chain reaction (PCR). The waxy gene of these genetic stocks had three types of length polymorphisms, suggesting that there
are insertion/deletion mutations at the 5′ leader sequence of the waxy gene. DNA sequence analysis of the polymorphic PCR
products showed that: (1) a 403-bp deletion mutation, which included a complete exon I, was found in the wax allele and a 193-bp insertion sequence was located in the intron I, and (2) the insertion sequence was also located in intron
I of the Wax allele. The identity of the insertion sequence was completely conserved between the wax allele and the novel Wax allele. These finding s implying that the wax allele, which was found in indigenous waxy barley, originated in non-waxy barley with the novel Wax allele.
Received: 12 January 2001 / Accepted: 17 April 2001 相似文献
3.
Zhen Wang Changbin Chen Yunyuan Xu Rongxi Jiang Ye Han Zhihong Xu Kang Chong 《Plant Molecular Biology Reporter》2004,22(4):409-417
In the last decade, RNA interferences (RNAi) has proven to be an effective strategy to knock out homologous genes in a wide
range of species. Based on its principle, a new generation of vectors containing an inverted target sequence separated by
an intron as a loop, developing simplifications to the procedure of RNAi construction are required to improve the efficiency
of gene inactivation techniques. Here, a novel polymerase chain reaction (PCR)—based RNAi vector pTCK303 with a maize ubiquitin
promoter, 2 specific multiple enzyme sites, and a rice intron was constructed for monocot gene silencing. With this vector,
only 1 PCR product amplified by a single pair of primers and 2 ligation reactions were needed to create an RNAi construct,
which shortened the time span before being transformed into the plant. To test the efficiency of vector pTCK303, a rice geneOsGAS1 was used, and its RNAi construct was introduced into rice calli. Southern blot analysis of the transgenic rice confirmed
the presence of theOsGAS1 RNAi structure. The decrease inOsGAS1 level in the transgenic rice was detected by Northern blot probed with anOsGAS1-specific sequence. Moreover, the rate of inhibition of the RNA expression level in RNAi transgenic rice was approximately
85% according to our real-time PCR. Therefore, the RNAi vector pTCK303 based on the homology-dependent gene-silencing mechanisms
facilitated the inhibition of endogenous genes in a monocot and was proven to be a practical and efficient platform for silencing
a rice gene.
These authors contributed equally to this work. 相似文献
4.
A new 5′ terminal murine GAPDH exon identified using 5′RACE LaNe 总被引:2,自引:0,他引:2
Park DJ 《Molecular biotechnology》2005,29(1):39-46
In this work, a ligation-independent, fully gene-specific, nested polymerase chain reaction (PCR) method for the elucidation
of 5′ cDNA sequence is described and demonstrated for the first time. Two manifestations of the method, rapid amplification
of cDNA ends (RACE) by lariat-dependent nested PCR 5′ (RACE LaNe), at least as simple to perform as conventional RACE, were
successfully applied to the murine housekeeping genes phosphoglycerate kinase 1 (PGK1), β-actin (β-ACT), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and the alpha thalassemia mental retardation Y homolog (ATRY) gene of the marsupial, Macropus eugenii. Significantly, a new murine GAPDH 5′ exon, separated by 365 kb of intronic sequence from previously annotated GAPDH sequence, was discovered using 5′RACE LaNe. 相似文献
5.
Gordonia amicalis F.5.25.8 has the unique ability to desulfurize dibenzothiophene and to metabolize carbazole [Santos et al., Appl Microbiol
Biotechnol 71:355–362, 2006]. Efforts to amplify the dsz genes from G. amicalis F.5.25.8 based on polymerase chain reaction (PCR) primers designed using the dsz gene sequences of Rhodococcus erythropolis IGTS8 were mostly unsuccessful. A comparison of the protein sequences of dissimilar desulfurization enzymes (DszABC, BdsABC,
and TdsABC) revealed multiple conserved regions. PCR primers targeting some of the most highly conserved regions of the desulfurization
genes allowed us to amplify dsz genes from G. amicalis F.5.25.8. DNA sequence data that include nearly the entirety of the desulfurization operon as well as the promoter region
were obtained. The most closely related dsz genes are those of G. alkinovorans strain 1B at 85% identity. The PCR primers reported here should be useful in microbial ecology studies and the amplification
of desulfurization genes from previously uncharacterized microbial cultures. 相似文献
6.
Inter-simple-sequence-repeat (ISSR) polymorphisms are useful for finding markers associated with disease resistance gene clusters 总被引:11,自引:0,他引:11
M. B. Ratnaparkhe M. Tekeoglu F. J. Muehlbauer 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,97(4):515-519
We describe a simple and new approach, based on inter-simple sequence repeats (ISSRs), for finding markers linked to clusters
of disease resistance genes. In this approach, simple sequence repeats (SSR) are used directly in PCR reactions, and markers
found to be linked to disease resistance genes provide important information for the selection of other sequences which can
be used with PCR to find other linked markers. Based on an ISSR marker linked to a gene of interest, many new markers can
be identified in the same region. We previously demonstrated that ISSR markers are useful in gene tagging and identified a
marker, UBC-855500, linked to the gene for resistance to fusarium wilt race 4 in chickpea. This ISSR marker provided the information used in
the present study for selecting other primers which amplified a region linked to the gene for resistance to fusarium wilt
race 4. The primers were based on homology with the (AC)n sequence and were used for PCR amplifications. Changes in the sequence were at the anchor region of the primers. The repeat
(AC)8T amplified a marker, UBC-8251200, which was located 5.0 cM from the gene for resistance to fusarium wilt race 4 and was closer than other markers. These results
indicated that ISSR markers can provide important information for the design of other primers and that by making changes at
the 3′ and 5′ anchors close linkage to the desired gene can be found. The approach allows rapid scanning of the targeted region
and may provide important information for genome analysis of plant species.
Received: 20 January 1998 / Accepted: 19 March 1998 相似文献
7.
Ayliffe MA Mitchell HJ Deuschle K Pryor AJ 《Molecular genetics and genomics : MGG》2005,274(5):494-505
Proline accumulation and catabolism play significant roles in adaptation to a variety of plant stresses including osmotic
stress, drought, temperature, freezing, UV irradiation, heavy metals and pathogen infection. In this study, the gene Δ1 -pyrroline-5-carboxylate dehydrogenase (P5CDH), which catalyzes the second step in the conversion of proline to glutamate, is characterized in a number of cereal species.
P5CDH genes from hexaploid wheat, Triticum turgidum (durum wheat), Aegilops tauschii, Triticum monococcum, barley, maize and rice were shown to be conserved in terms of gene structure and sequence, present as a single copy per
haploid, non-polyploid genome and located in evolutionarily conserved linkage groups. A wheat cDNA sequence was shown by yeast
complementation to encode a functional P5CDH activity. A divergently-transcribed rab7 gene was identified immediately 5′ of P5CDH in all grasses examined, except rice. The rab7/P5CDH intergenic region in these species, which presumably encompasses 5′ regulatory elements of both genes, showed a distinct
pattern of sequence evolution with sequences in juxtaposition to each ORF conserved between barley, wheat, A. tauschii and T. monococcum. More distal 5′ sequence in this intergenic region showed a higher rate of divergence, with no homology observed between
these regions in the wheat and barley genomes. Maize and rice showed no similarity in regions 5′ of P5CDH when compared with wheat, barley, and each other, apart from a 22 bp region of conserved non-coding sequence (CNS) that is
similar to a proline response element identified in the promoter of the Arabidopsis proline dehydrogenase gene. A palindromic
motif similar to this cereal CNS was also identified 5′ of the Arabidopsis AtP5CDH gene showing conservation of this sequence in monocot and dicot lineages. 相似文献
8.
9.
Jian Liu Tingzhao Rong Wanchen Li 《Molecular breeding : new strategies in plant improvement》2007,20(2):93-102
Four pairs of specific PCR primers have been designed on the basis of the sequence of the granule-bound starch synthase gene
(GBSS; dominant non-waxy gene Wx) and used to amplify its homologous sequence from thirteen waxy and two non-waxy inbred lines. Results from electrophoresis
indicated that the recessive waxy gene was wx, derived from the dominant non-waxy gene Wx by mutation at its 3′ end. The sequence of the mutated 3′ end was amplified by the TAIL-PCR technique. Sequence alignment
showed that the mutation of the wx gene was caused by transposition of the aldehyde dehydrogenase gene rf2. Two pairs of specific primers were designed on the basis of the sequence difference between the dominant gene Wx and its mutated recessive allele wx and used as intragenic selection markers to identify individual plants of genotypes WxWx, Wxwx, and wxwx by PCR amplification from the segregating population of the F2 generation crossed between waxy and non-waxy inbred lines. Iodine solution staining and starch component assay showed that
all the 35 F2 plants identified as genotype WxWx produced non-waxy kernels of the F3 generation and that all 33 F2 plants identified as genotype wxwx produced waxy kernels of the F3 generation. This result can be used to improve the selection efficiency of waxy maize breeding and for selection of other
single genes and major polygenes. 相似文献
10.
Resistance gene analogues from rice: cloning, sequencing and mapping 总被引:18,自引:0,他引:18
R. Mago S. Nair M. Mohan 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,99(1-2):50-57
Degenerate oligonucleotide primers were designed on the basis of nucleotide-binding-site (NBS) motifs conserved between resistance
genes of Arabidopsis, flax and tobacco and subsequently used as PCR primers to amplify resistance gene analogues (RGA) in rice. Primers amplified
a major band of approximately 500 bp. Restriction analysis of the amplified product revealed that the band was made up of
several different fragments. Many of these fragments were cloned. Sixty different cloned fragments were analysed and assigned
to 14 categories based on Southern blot analysis. Fourteen clones, each representing one of the 14 categories of RGAs were
mapped onto the rice genetic map using a Nipponbare ( japonica)בKasalath’ (indica) mapping population consisting of 186 F2 lines. Of the 14 clones representing each class 12 could be mapped onto five different chromosomes of rice with a major cluster
of 8 RGAs on chromosome 11. Our results indicate that it is possible to use sequence homology from conserved motifs of known
resistance genes to amplify candidate resistance genes from diverse plant taxa.
Received: 23 September 1998 / Accepted: 28 November 1998 相似文献
11.
Genome scanning for resistance-gene analogs in rice, barley, and wheat by high-resolution electrophoresis 总被引:31,自引:8,他引:23
X. M. Chen R. F. Line H. Leung 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,97(3):345-355
Genes cloned from diverse plants for resistance to different pathogens have sequence similarities in domains presumably involved
in pathogen recognition and signal transduction in triggering the defense response. Primers based on the conserved regions
of resistance genes often amplify multiple fragments that may not be separable in an agarose gel. We used denaturing polyacrylamide-gel
electrophoresis to detect PCR products of plant genomic DNA amplified with primers based on conserved regions of resistance
genes. Depending upon the primer pairs used, 30–130 bands were detected in wheat, rice, and barley. As high as 47%, 40%, and
27% of the polymorphic bands were detected in rice, barley, and wheat, respectively, and as high as 12.5% of the polymorphic
bands were detected by certain primers in progeny from a cross of the wheat cultivars ‘Stephens’ and ‘Michigan Amber’. Using
F6 recombinant inbred lines from the ‘Stephens’בMichigan Amber’ cross, we demonstrated that polymorphic bands amplified with
primers based on leucine-rich repeats, nucleotide-binding sites and protein kinase genes, were inherited as single loci. Linkages
between molecular markers and stripe rust resistance genes were detected. This technique provides a new way to develop molecular
markers for assessing the genetic diversity of germplasm based upon potential candidate resistance genes in diverse species.
Received : 5 September 1997 / Accepted : 6 November 1997 相似文献
12.
PCR with degenerate primers amplifies a subgenomic DNA fragment from the endoglucanase gene(s) of Torula thermophila, a thermophilic fungus 总被引:1,自引:0,他引:1
The aim of this study was to enable the polymerase chain reaction (PCR) amplification of DNA fragments within endoglucanase
gene(s) of Torula thermophila, by using degenerate primers so that the amplified fragment(s) could be used as homologous probe(s) for cloning of full-length
endoglucanase gene(s). The design of the degenerate PCR primers was mainly based on the endoglucanase sequences of other fungi.
The endoglucanase gene sequence of Humicola insolens was the only sequence from a thermophilic fungus publicly available in the literature. Therefore, the endoglucanase sequences
of the two Trichoderma species, Trichoderma reesei and Trichoderma longibrachiatum, were used to generalize the primers. PCR amplification of T. thermophila genomic DNA with these primers resilied in a specific amplification. The specificity of the amplified fragment was shown
by Southern hybridization analysis using egl3 gene of T. reesei as probe. This result suggested that the degenerate primers used in this study may be of value for studies aimed at cloning
of endoglucanase genes from a range of related fungi. 相似文献
13.
Chan Sun Park 《Journal of applied phycology》2006,18(2):203-207
The fungal parasite Pythium porphyrae is the causative organism of red rot disease in Porphyra cultivation farms. The detection of P. porphyrae from dried Porphyra yezoensis sheets was achieved using the species-specific primers PP-1 (5′-TGTGTTCTGTGCT-CCTCTCG-3′) and PP-2 (5′-CCCAAATTGGTGTTGCCTCC-3′) with the polymerase chain reaction (PCR). The DNA sequence (707 bp) of PCR product was found to be identical to that amplified from ITS rDNA extracted from a type species of P. porphyrae (IFO 30800, The Institute of Fermentation, Osaka, Japan). Quantities of the product amplified varied with the time when samples were harvested after the occurrence of red rot disease in Porphyra farms. This simple, rapid, and inexpensive method should have great applications in furthering quality control and determination of quality ranking in the Porphyra processing industry. 相似文献
14.
Diversity in the Integrase Coding Domain of a gypsy-like Retrotransposon Among Wild Relatives of Rice in the Oryza officinalis Complex 总被引:1,自引:0,他引:1
The Oryza officinalis complex is a genetically diverse, tertiary genepool of rice. We analyzed part of the primary structure of the integrase coding
domain (ICD) of a gypsy-like retrotransposon from species of the O. officinalis species complex. PCR was performed with degenerate primers that hybridized to conserved sequences in the integrase genes
of gypsy-type retrotransposons, using total DNA from different species of the O. officinalis complex as templates. Cloning and sequencing of the PCR products showed that the amplified fragments are highly homologous
to each other (75–90%) and belong to one family of retrotransposons that is related to the previously studied RIRE-2 element from rice. Two main subfamilies of 292 and 351 bp were distinguished. Analysis of primary sequence data supports
previous reports that sequence divergence during vertical transmission has been the major influence on the evolution of gypsy-type retrotransposons in Oryza species. Based on sequence data phylogenetic relationships among species of the O. officinalis complex were estimated. The data suggests that O. eichingeri is more closely related to the ancestral species of the complex.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
15.
A. I. Martsinkovskaya R. S. Moukhamedov A. A. Abdukarimov 《Plant Molecular Biology Reporter》1996,14(1):44-49
Our purpose was to develop a new approach to the identification ofGossypium cotton varieties and species based on polymerase chain reaction (PCR). Species-specific distinctions within the genusGossypium have been detected by the amplification of ribosomal genes, namely theRrn18-Rrn25 internal transcribed spacer (ITS) regions that had sequence differences. Using the primers to the 3′-end ofRrn18 adjacent to ITS1 and the 5′-end ofRrn25 adjacent to ITS2 from tomato, we have obtained amplified fragments of two cotton species,G. barbadense andG. herbaceum. Interspecies distinctions have been revealed by the restriction assay of these amplification products. The restriction patterns
are distinguished not only by number but by location and intensity of the bands. Our results illustrate the effective use
of differences in ribosomal intergenic sequences for the differentiation of varieties and species ofGossypium. 相似文献
16.
Differential display (DD) is one of the most commonly used approaches for identifying differentially expressed genes. However,
there has been lack of an accurate guidance on how many DD polymerase chain reaction (PCR) primer combinations are needed
to display most of the genes expressed in a eukaryotic cell. This study critically evaluated the gene coverage by DD as a
function of the number of arbitrary primers, the number of 3′ bases of an arbitrary primer required to completely match an
mRNA target sequence, the additional 5′ base match(s) of arbitrary primers in first-strand cDNA recognition, and the length
of mRNA tails being analyzed. The resulting new DD mathematical model predicts that 80 to 160 arbitrary 13mers, when used
in combinations with 3 one-base anchored oligo-dT primers, would allow any given mRNA within a eukaryotic cell to be detected
with a 74% to 93% probability, respectively. The prediction was supported by both computer simulation of the DD process and
experimental data from a comprehensive fluorescent DD screening for target genes of tumor-suppressor p53. Thus, this work provides a theoretical foundation upon which global analysis of gene expression by DD can be pursued. 相似文献
17.
Differential expression of manganese superoxide dismutase sequence variants in near isogenic lines of wheat during cold acclimation 总被引:7,自引:0,他引:7
Numerous sequence variants of wheat (Triticum aestivum L.) manganese superoxide dismutase (MnSOD) genes have been found. Quantitative real-time PCR was used to measure the expression
levels of three MnSOD genes distinguished by a variable amino acid, and three genes distinguished by sequence variation in
the 3′ untranslated region (3′ UTR), in wheat plants grown at 20°C and cold-acclimated for 1–4 weeks at 2°C. The amino acid
variants did not differ significantly in expression levels, however, differential expression of genes differing in the 3′
UTR was observed. Diploid wheat-related species also carried sequence variants of MnSOD, with differing levels of expression,
suggesting diversification of the MnSOD gene family occurred prior to the polyploidization events of hexaploid wheat. 相似文献
18.
Development of microsatellite markers from tartary buckwheat 总被引:2,自引:0,他引:2
A genomic library enriched with (gT)n repeats from tartary buckwheat (Fagopyrum tataricum) was constructed using 5′-anchored PCR for the development of microsattellite markers. Sequencing analysis of 5 clones from
the library showed that they all contained microsatellites (totally 10 loci), and each was unique. An additional locus-specific
primer was designed according to flanking sequence. Two of the microsatellite loci of 10 tartary buckwheat varieties were
amplified using an anchored primer and a locus-specific primer, which revealed a clear polymorphic pattern. The data confirmed
that the degenerate primer was reliably anchoring at the 5′-end of the microsatellite, and the primers developed based on
this technology could be used for diversity analysis of tartary buckwheat. 相似文献
19.
Fine Mapping and in silico Isolation of the EUI1 Gene Controlling Upper Internode Elongation in Rice 总被引:8,自引:0,他引:8
Upper internode elongation in rice is an important agronomic trait. Well-known mutants with an elongated uppermost internode
(eui) are important germplasms for developing unsheathed-panicle male-sterile lines in hybrid rice breeding. We finely mapped
the eui1 gene and identified its candidate gene using in silico analysis based on previous research work and rice genomic sequence data. The rice eui1 gene was mapped to two overlapping BAC clones, OSJNBa0095J22 and OSJNBb0099O15, between the markers AC40 and AC46, that were
0.64 cM apart and spanned approximately 152 kb. A simple sequence repeat (SSR) marker AC41 that cosegregated with eui1 was located in an intron of a putative cytochrome P450-related gene. In silico analysis suggested that this encoded the cytochrome CYP714D1. Allelic sequencing confirmed that EUI1 corresponded to this P450 gene. A gamma ray-induced eui1 mutant carried a deletion in exon II of the EUI1 gene, and resulted in a frame-shift deletion that produced a truncated polypeptide. We conclude that the EUI1 gene controlling the upper internode elongation in rice is 9804 bp long, and comprises two exons and one intron. The length
of the cDNA is 1931 bp containing a 1734 bp ORF, a 110 bp 5′-UTR and a 87 bp 3′-UTR. The ORF encodes an unknown 577 amino
acid functional protein, that appears to be a member of the cytochrome P450 family.
Hongli Ma, Shubiao Zhang: These authors contributed equally to this work 相似文献
20.
Target region amplification polymorphism: A novel marker technique for plant genotyping 总被引:51,自引:0,他引:51
The advent of large-scale DNA sequencing technology has generated a tremendous amount of sequence information for many important
organisms. We have developed a rapid and efficient PCR-based technique, which uses bioinformatics tools and expressed sequence
tag (EST) database information to generate polymorphic markers around targeted candidate gene sequences. This target region
amplification polymorphism (TRAP) technique uses 2 primers of 18 nucleotides to generate markers. One of the primers, the
fixed primer, is designed from the targeted EST sequence in the database; the second primer, the arbitrary primer, is an arbitrary
sequence with either an AT-or GC-rich core to anneal with an intron or exon, respectively. PCR amplification is run for the
first 5 cycles with an annealing temperature of 35°C, followed by 35 cycles with an annealing temperature of 50°C. For different
plant species, each PCR reaction can generate as many as 50 scorable fragments with sizes ranging from 50–900 bp when separated
on a 6.5% polyacrylamide sequencing gel. The TRAP technique should be useful in genotyping germplasm collections and in tagging
genes governing desirable agronomic traits of crop plants. 相似文献