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Results for “spatial gene expression”

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PubMed2000

Differentiation and histological analysis of embryonic stem cell-derived neural transplants in mice.

We report here that neural transplantation of in vitro-differentiated embryonic stem (ES) cells provides a versatile strategy for gene transfer into the central nervous system. ES cells were subjected to an optimized in vitro differentiation protocol to obtain embryoid bodies. These aggregates were stereotaxically transplanted into the brain of recipient adult mice, where they followed a strictly controlled differentiation pattern and eventually formed mature neural grafts. A marker gene, introduced into the ROSA26 locus allowed for precise determination of the fate of the descendants of the transplanted embryoid bodies and revealed that not only neurons but also astrocytes, oligodendrocytes and even microglial cells were graft-derived. Evaluation of long-term experiments showed viable grafts with a stable transgene expression and proved that this approach provides a tool for reliable gene expression within a spatially delimited area of neural tissue.

Animals↗
PubMed1997

Mechanisms of evolutionary changes in timing, spatial expression, and mRNA processing in the msp130 gene in a direct-developing sea urchin, Heliocidaris erythrogramma.

Developmental processes associated with skeletogenesis differ in the direct-developing sea urchin Heliocidaris erythrogramma from that in Heliocidaris tuberculata and other indirect-developing species. In H. erythrogramma, the differences include ingression of a much higher number of mesenchyme cells, failure of the cells to form the typical ring pattern of cells prior to the onset of skeletogenesis, a significantly reduced larval skeleton, and a delay in timing of expression of the skeletogenic cell-restricted gene msp130. We report that the heterochronic change in msp130 expression is regulated at the level of transcription. By transient expression of reporter constructs containing msp130 promoter regions from direct- and indirect-developing species, we found that this evolutionary change in regulation is consistent with changes in the timing of action of trans-acting factors in skeletogenic mesenchyme cells. We further used these experiments to show that the H. erythrogramma promoter contains elements required for correct spatial expression in the primary mesenchyme cells of an indirect-developing host. We finally show that alternate processing of H. erythrogramma msp130 is thus far specific to this species and not an aspect of adult skeletogenesis.

Alternative Splicing↗
PubMed2004

Direct visualization of electroporation-assisted in vivo gene delivery to tumors using intravital microscopy - spatial and time dependent distribution.

BACKGROUND: Electroporation is currently receiving much attention as a way to increase drug and DNA delivery. Recent studies demonstrated the feasibility of electrogene therapy using a range of therapeutic genes for the treatment of experimental tumors. However, the transfection efficiency of electroporation-assisted DNA delivery is still low compared to viral methods and there is a clear need to optimize this approach. In order to optimize treatment, knowledge about spatial and time dependency of gene expression following delivery is of utmost importance in order to improve gene delivery. Intravital microscopy of tumors growing in dorsal skin fold window chambers is a useful method for monitoring gene transfection, since it allows non-invasive dynamic monitoring of gene expression in tumors in a live animal. METHODS: Intravital microscopy was used to monitor real time spatial distribution of the green fluorescent protein (GFP) and time dependence of transfection efficiency in syngeneic P22 rat tumor model. DNA alone, liposome-DNA complexes and electroporation-assisted DNA delivery using two different sets of electric pulse parameters were compared. RESULTS: Electroporation-assisted DNA delivery using 8 pulses, 600 V/cm, 5 ms, 1 Hz was superior to other methods and resulted in 22% increase in fluorescence intensity in the tumors up to 6 days post-transfection, compared to the non-transfected area in granulation tissue. Functional GFP was detected within 5 h after transfection. Cells expressing GFP were detected throughout the tumor, but not in the surrounding tissue that was not exposed to electric pulses. CONCLUSIONS: Intravital microscopy was demonstrated to be a suitable method for monitoring time and spatial distribution of gene expression in experimental tumors and provided evidence that electroporation-assisted gene delivery using 8 pulses, 600 V/cm, 5 ms, 1 Hz is an effective method, resulting in early onset and homogenous distribution of gene expression in the syngeneic P22 rat tumor model.

Animals↗
PubMed1998

Effects of mutation of the Olf-1 motif on transgene expression in olfactory receptor neurons.

We have examined the effect of mutating the Olf-1 binding motif of the olfactory marker protein (OMP) promoter in determining olfactory neuron-specific gene expression in adult tissues and during embryonic development. The proximal Olf-1 motif located 170 nucleotides upstream of the transcription start site of the OMP gene was mutated to prevent its interaction with the Olf-1 factor in vitro. The wild-type and mutated fragments of the OMP gene extending from -239 to +55 nucleotides relative to the transcription start site were used to direct expression of a lacZ reporter gene in transgenic mice. The transgenic animals were analyzed for cell-specific and developmental expression of the reporter gene. We demonstrate that the mutation that prevents interaction of Olf-1 with its binding site does not alter the temporal and spatial patterns of gene expression in olfactory sensory neurons but does alter the specificity and level of expression in other neuronal populations. These observations are consistent with our demonstration that the mutated Olf-1 site interacts with nuclear proteins present in the central nervous system (CNS).

Animals↗
PubMed2002

Screen for stage-specific expression genes between tail bud stage and heartbeat beginning stage in embryogenesis of gynogenetic silver crucian carp.

A systemic study was initiated to identify stage-specific expression genes in fish embryogenesis by using suppression subtractive hybridization (SSH) technique. In this study, we presented a preliminary result on screen for stage-specific expression genes between tail bud stage (TBS) and heartbeat beginning stage (HBS) in gynogenetic silver crucian carp (Carassius auratus gibelio). Two SSH plasmid libraries specific for TBS embryos and HBS embryos were constructed, and stage-specific expression genes were screened between the two stages. 1963 TBS positive clones and 2466 HBS positive clones were sampled to PCR amplification, and 1373 TBS and 1809 HBS PCR positive clones were selected to carry out dot blots. 169 TBS dot blot positive clones and 272 HBS dot blot positive clones were sequenced. Searching GenBank by using these nucleotide sequences indicated that most of the TBS dot blot positive clones could not be found homologous sequences in the database, while known genes were mainly detected from HBS dot blot positive clones. Of the 79 known genes, 20 were enzymes or kinases involved in important metabolism of embryonic development. Moreover, specific expressions of partial genes were further confirmed by virtual northern blots. This study is the first step for making a large attempt to study temporal and spatial control of gene expression in the gynogenetic fish embryogenesis.

Animals↗
PubMed2002

ICSBP/IRF-8 transactivation: a tale of protein-protein interaction.

Interferon (IFN) consensus sequence binding protein (ICSBP) is a member of a family of transcription factors termed IFN regulatory factors (IRF) and is also called IRF-8. Its expression is restricted mainly to cells of the immune system, and it plays a key role in the maturation of macrophages. ICSBP exerts its activity through the formation of different DNA-binding heterocomplexes. The interacting partner dictates a specific DNA recognition sequence, thus rendering ICSBP dual transcriptional activity, that is, repression or activation. Accordingly, such DNA elements were identified at the promoter regions of target genes that manifest macrophage action. A specific module (IRF association domain [IAD]) within ICSBP and a PEST domain located on the interacting partners mediate this association. Thus, ICSBP serves as an excellent prototype, demonstrating how a small subset of transcription factors can regulate gene expression in a spatial, temporal, and delicate tuning through combinatorial protein-protein interactions on different enhanceasomes.

Animals↗
PubMed2007

Expression of steroid receptor coactivator-1 is elevated during neuronal differentiation of murine neural stem cells.

The cell-lineage-specific fate of neural stem cells (NSCs) is defined by spatial and temporal gene expression regulated by various transcription factors and their coregulators, including coactivators and corepressors. To clarify the cellular distribution of the steroid receptor coactivator-1 (SRC-1), a ligand-dependent nuclear receptor coactivator, during neurogenesis, we examined the expression profiles of SRC-1 during the proliferation and differentiation in culture of NSCs derived from the ganglionic eminence of mouse embryos. We found that SRC-1 was rarely expressed in proliferating cells and multipotent precursors with the typical characters of NSCs. Under conditions that promote the differentiation of NSCs, both the SRC-1 transcript and protein levels were elevated and the population of SRC-1-positive cells was found to be higher in the mature neurons than in the immature neurons. In contrast, SRC-1 expression was rarely localized in the glial lineage cells, including astrocytes and oligodendrocytes. These results indicate that SRC-1 expression accompanies the appearance of neuronal-fate-committed cells derived from multipotent NSCs and is preferentially expressed in neuronal lineage cells during the differentiation of NSCs.

Analysis of Variance↗
PubMed1995

Functional involvement of Pax-1 in somite development: somite dysmorphogenesis in chick embryos treated with Pax-1 paired-box antisense oligodeoxynucleotide.

The metameric pattern of the vertebrate axial skeleton, defined by structures such as the vertebral bodies and ribs, is a result of segmentation events that occur during embryogenesis. The key event in axial segmentation is somite formation. This study examines the role of Pax-1, a member of the paired-box containing Pax gene family, in chick somite development. To investigate whether misexpression of Pax-1 during somite development is functionally related to abnormal axial patterning, antisense methodologies were used to perturb Pax-1 expression. An antisense, phosphorothioate-modified oligodeoxynucleotide (ODN) was designed based on the mouse Pax-1 paired-box sequence, and was either injected into or directly applied topically to early, somitic stage chick embryos. Abnormalities in somite structure and pattern were subsequently observed and scored, including loss of somites (18% of injected embryos, 35% of embryos treated by topical application), fusion of somites (25% of injected, 6% with topical application), and shortened body axis (0% of injected, 11% with topical application). Control embryos receiving sense ODN or non-sense ODN (a scrambled sequence with base composition identical to the antisense ODN) showed substantially fewer somite anomalies, indicating that the effects were sequence-specific. These developmental abnormalities were analyzed using standard histological methods. Whole mount in situ hybridization was carried out to analyze the three-dimensional pattern of Pax-1 expression in whole embryos. In control, untreated embryos, the expression was localized to the entire epithelial somite, and as the somite matured, the expression was limited to its ventromedial region. With Pax-1 antisense ODN treatment, embryos with fused somites retained expression over the entire fused somite, and embryos that had complete loss of somites had greatly reduced expression of Pax-1 transcript. The results presented here provide strong evidence that Pax-1 is functionally important during somitogenesis and morphogenesis of the vertebral column. The spatial pattern of gene expression appears to delineate different populations of cells in the developing embryo (i.e., somite from somite, sclerotome from dermomyotome), and is consistent with the hypothesis that Pax-1 is involved in forming or maintaining boundaries at specific times and locations during development.

Abnormalities, Drug-Induced↗
PubMed1999

Interaction between Notch signalling and Lunatic fringe during somite boundary formation in the mouse.

BACKGROUND: The process of somitogenesis can be divided into three major events: the prepatterning of the mesoderm; the formation of boundaries between the prospective somites; and the cellular differentiation of the somites. Expression and functional studies have demonstrated the involvement of the murine Notch pathway in somitogenesis, although its precise role in this process is not yet well understood. We examined the effect of mutations in the Notch pathway elements Delta like 1 (Dll1), Notch1 and RBPJkappa on genes expressed in the presomitic mesoderm (PSM) and have defined the spatial relationships of Notch pathway gene expression in this region. RESULTS: We have shown that expression of Notch pathway genes in the PSM overlaps in the region where the boundary between the posterior and anterior halves of two consecutive somites will form. The Dll1, Notch1 and RBPJkappa mutations disrupt the expression of Lunatic fringe (L-fng), Jagged1, Mesp1, Mesp2 and Hes5 in the PSM. Furthermore, expression of EphA4, mCer 1 and uncx4.1, markers for the anterior-posterior subdivisions of the somites, is down-regulated to different extents in Notch pathway mutants, indicating a global alteration of pattern in the PSM. CONCLUSIONS: We propose a model for the mechanism of somite border formation in which the activity of Notch in the PSM is restricted by L-fng to a boundary-forming territory in the posterior half of the prospective somite. In this region, Notch function activates a set of genes that are involved in boundary formation and anterior-posterior somite identity.

Animals↗
PubMed2000

Modulation in the developmental expression profile of Sp1 subsequent to transplacental exposure of fetal rats to desorbed benzo[a]pyrene following maternal inhalation.

Any alteration of the critical sequence of genes that are required to coordinate the differentiation of cells, the promotion of migration, dendritic arborization, synapse formation, and myelination in the developing nervous system would be expected to have deleterious consequences. The focus of this article is a molecular evaluation of the neurotoxicological effects that result subsequent to the transplacental exposure of fetal rats to desorbed benzo(a)pyrene (BaP) following maternal inhalation. A state-of-the-art, newly designed, fabricated, and tested model aerosol generation system was utilized in these studies. Timed-pregnant Sprague Dawley rats were exposed for 4 h on gestation day 15 of a 21-day gestation period to an acute dose of BaP:carbon black aerosol (100 microg/m(3)). Controls received carbon black only. Nominal and chamber concentrations of the particulate aerosol were determined gravimetrically with a seven-stage cascade impactor. The aerosol exhibited a trimodal distribution with 95% cumulative mass less than 15.85 microm, 90% cumulative mass less than 10 microm, 67. 5% cumulative mass less than 2.5 microm and 66.2% cumulative mass less than 1.0 microm. Time-course bioavailability results indicated that greater than 95% of the parent compound is cleared from blood 240 min postexposure. An Sp1 transcription factor consensus sequence was examined by electrophoretic mobility shift analysis of nuclear extracts from various brain regions of resulting pups on postnatal days 3, 5, 7, 10, and 15. It revealed perturbations in the developmental expression profile of Sp1 abundance as a result of nose-only particulate aerosol exposure to the timed-pregnant dam. The data obtained on the temporal and spatial regulation of gene expression in the brain indicate that (1) Sp1 DNA-binding is developmentally regulated and expressed very highly in actively developing brain regions, and (2) a consequence of the transplacental deposition of desorbed BaP to the fetus is in utero neurotoxicity.

Administration, Inhalation↗
PubMed2003

Potato MADS-box gene POTM1-1 transcripts are temporally and spatially distributed in floral organs and vegetative meristems.

The POTM1-1 gene is abundantly expressed in both vegetative and reproductive organs of potato. We performed in situ hybridization and RNA blotting analysis to investigate the patterns of POTM1-1 gene expression in the flower development and the early tuber development. In the early flowers, POTM1-1 transcripts were accumulated abundantly in the developing reproductive organs, including the placentae of carpels and the pollen sacs of stamens. In contrast, the pattern of POTM1-1 distribution during late flower development was different from that of the early flower development. The POTM1-1 transcripts were abundant in the sepals and petals of late flowers, but were minimally expressed in the stamens and carpel. In the shoot apical meristem of the vegetative organs, transcripts were distributed throughout meristem domes, young leaves, and developing vascular cambium. In the early tuberization, the transcripts were widely distributed in the swollen tips of the stolons. Taken together, the results suggest that POTM1-1 gene expression is temporally and spatially regulated in active growing tissues of both vegetative and floral organs with specific distribution patterns dependent upon the developmental stages of the tissue.

Flowers↗
PubMed1999

Molecular characterization of Zfp54, a zinc-finger-containing gene that is deleted in the embryonic lethal mutation tw18.

Mapping studies have indicated that hundreds of Kruppel-type zinc-finger (ZNF)-containing genes are present within the genomes of mammals. One-third or more of these genes contain a second, highly conserved element termed the Kruppel-associated box (KRAB). In a recent study, several partial cDNA clones encoding distinct KRAB elements were mapped in mouse. One of these sequences, clone KRAB10, was assigned to mouse Chromosome (Chr) 9. We have isolated the complete coding portion of the KRAB10-containing gene and demonstrate that it is identical to Zfp54, one of several related ZNF genes that are located on mouse Chr 17 in the interval that is deleted in the tw18 mutation. Sequence analysis has shown that the KRAB A domain encoded by Zfp54 is highly similar to the comparable motif encoded by its nearest known neighbor, Zfp51. However, other portions of the related proteins, including their ZNF domains, are noticeably different in structure. Studies of Zfp54 expression revealed the presence of transcript in several adult tissues and in embryos throughout development; however, elevated levels of transcript were detected only in adult testis and in 7-day-old embryos. These data suggest that Zfp54 gene expression is under spatial and temporal control and may, therefore, play important roles in male germ cell and embryonic development.

Animals↗
PubMed2026

Machine learning and multi-omics clustering to map cellular rewiring and immune evasion in ccRCC.

Immune checkpoint blockade (ICB) efficacy in clear cell renal cell carcinoma (ccRCC) is limited by tumor microenvironment (TME) heterogeneity. Because traditional bulk-derived models lack spatial resolution, we developed an integrated framework connecting macroscopic survival risks to microscopic TME structures. We applied ten algorithms to establish multi-omics subtypes and evaluated 101 machine-learning combinations across three independent cohorts to generate a Consensus Machine Learning-driven Signature (CMLS). The signature's spatial and cellular origins were decoded using spatial transcriptomics (ST) and a 140,000-cell scRNA-seq atlas. Expression of key genes was experimentally validated via RT-qPCR in 17 paired ccRCC clinical tissues. We identified two molecular subtypes with distinct clinical and epigenetic profiles. SuperPC optimization yielded a 24-gene CMLS serving as an independent prognostic factor. scRNA-seq and ST deconvolution revealed these signals predominantly originate from cancer-associated fibroblasts (CAFs) and malignant epithelial cells, which collaborate to drive spatial immune exclusion. RT-qPCR confirmed significant overexpression of five core CMLS genes in ccRCC versus adjacent normal tissues. Low CMLS scores correlated with enhanced ICB responsiveness, whereas high-CMLS tumors demonstrated specific vulnerability to dasatinib and dabrafenib. The CMLS translates spatial immune-exclusion dynamics into a quantifiable metric, outperforming tumor mutational burden in predicting ICB benefits, providing a robust tool for patient stratification in ccRCC.

Humans↗
PubMedDate not supplied

Spatio-temporal characterization of retinal opsin gene expression during thyroid hormone-induced and natural development of rainbow trout.

The abundance and spatial distribution of retinal cone photoreceptors change during thyroid hormone (TH)-induced and natural development of rainbow trout (Oncorhynchus mykiss). These changes are thought to allow the fish to adapt to different photic environments throughout its life history. To date, the ontogeny of rainbow trout cone photoreceptors has been examined using physiological and morphological approaches. In this study, we extended these observations by measuring opsin gene expression in retinal quadrants during natural and TH-induced development. Gene expression during natural development was investigated in retinae from fish at both parr and smolt stages. The role of TH in modulating opsin gene expression was determined in TH-treated parr and control fish sampled after two, nine, and 22 days of treatment. Total RNA was isolated from each retinal quadrant and steady-state opsin mRNA levels were measured using reverse transcriptase real-time quantitative polymerase chain reaction (QPCR) analysis. Expression of ultraviolet-sensitive opsin (SWS1), rod opsin (RH1), middle wavelength-sensitive opsin (RH2), and long wavelength-sensitive opsin (LWS) transcripts vary spatially in the parr retina. Smolts, compared to parr, had downregulated SWS1 expression in all quadrants, lower LWS expression dorsally, higher RH1 expression nasally, and higher RH2 expression dorsally. In TH-treated parr, SWS1 opsin expression was downregulated in the nasal quadrants by two days. SWS1 displayed the greatest degree of downregulation in all quadrants after nine days of treatment, with an increase in short wavelength-sensitive (SWS2) and RH2 opsin mRNA expression in the temporal quadrants. This study reveals that opsin genes display spatially significant differences within rainbow trout retina in their level of mRNA expression, and that regulation of opsin expression is a dynamic process that is influenced by TH. This is particularly evident for SWS1 gene expression in parr following TH-induced and natural development.

Animals↗
PubMed1995

A 20-nucleotide element in the intestinal fatty acid binding protein gene modulates its cell lineage-specific, differentiation-dependent, and cephalocaudal patterns of expression in transgenic mice.

A sequence of epithelial cell proliferation, allocation to four principal lineages, migration-associated differentiation, and cell loss occurs along the crypt-villus axis of the mouse intestine. The sequence is completed in a few days and is recapitulated throughout the life-span of the animal. We have used an intestine-specific fatty acid binding protein gene, Fabpi, as a model for studying regulation of gene expression in this unique developmental system. Promoter mapping studies in transgenic mice identified a 20-bp cis-acting element (5'-AGGTGGAAGCCATCACACTT-3') that binds small intestinal nuclear proteins and participates in the control of Fabpi's cephalocaudal, differentiation-dependent, and cell lineage-specific patterns of expression. Immunocytochemical studies using confocal and electron microscopy indicate that it does so by acting as a suppressor of gene expression in the distal small intestine/colon, as a suppressor of gene activation in proliferating and nonproliferating cells located in the crypts of Lieberkühn, and as a suppressor of expression in the growth factor and defensin-producing Paneth cell lineage. The 20-bp domain has no obvious sequence similarities to known transcription factor binding sites. The three functions modulated by this compact element represent the types of functions required to establish and maintain the intestine's remarkably complex spatial patterns of gene expression. The transgenes described in this report also appear to be useful in characterizing the crypt's stem cell hierarchy.

Animals↗
PubMed2004

A leader intron and 115-bp promoter region necessary for expression of the carnation S-adenosylmethionine decarboxylase gene in the pollen of transgenic tobacco.

The expression of CSDC9 encoding S-adenosylmethionine decarboxylase (SAMDC) is developmentally and spatially regulated in carnation. To examine the regulation of the SAMDC gene, we analyzed the spatial expression of CSDC9 with a 5'-flanking beta-glucuronidase fusion in transgenic tobacco plants. GUS was strongly expressed in flower, pollen, stem and vein of cotyledons. Expression in both anther and stigma was under developmental control; analysis of a series of mutants with deletions of the 5'-flanking region demonstrated differential activation in petal, anther, stigma and pollen grains. All the major cis-regulatory elements required for pollen-specific transcription were located in the upstream region between -273 and -158. This region contains four putative elements related to gibberellin induction (pyrimidine boxes, TTTTTTCC and CCTTTT) and pollen-specific expression (GTGA and AGAAA). In addition, the first 5'-leader intron was necessary for tissue-specific expression.

5' Flanking Region↗
PubMed1992

Spatial and temporal expression pattern during sea urchin embryogenesis of a gene coding for a protease homologous to the human protein BMP-1 and to the product of the Drosophila dorsal-ventral patterning gene tolloid.

A cDNA clone coding for a sea urchin embryonic protein was isolated from a prehatching blastula lambda gt11 library. The predicted translation product is a secreted 64 x 10(3) Mr enzyme designated as BP10. The protein contains several domains: a signal peptide, a putative propeptide, a catalytic domain with an active center typical of a Zn(2+)-metalloprotease, an EGF-like domain and two internal repeats similar to repeated domains found in the C1s and C1r serine proteases of the complement cascade. The BP10 protease is constructed with the same domains as the human bone morphogenetic protein BMP-1, a protease described as a factor involved in bone formation, and as the recently characterized product of the tolloid gene which is required for correct dorsal-ventral patterning of the Drosophila embryo. The transcription of the BP10 gene is transiently activated around the 16- to 32-cell stage and the accumulation of BP10 transcripts is limited to a short period at the blastula stage. By in situ hybridization with digoxygenin-labelled RNA probes, the BP10 transcripts were only detected in a limited area of the blastula, showing that the transcription of the BP10 gene is also spatially controlled. Antibodies directed against a fusion protein were used to detect the BP10 protein in embryonic extracts. The protein is first detected in early blastula stages, its level peaks in late cleavage, declines abruptly before ingression of primary mesenchyme cells and remains constant in late development. The distribution of the BP10 protein during its synthesis and secretion was analysed by immunostaining blastula-stage embryos. The intracellular localization of the BP10 staining varies with time. The protein is first detected in a perinuclear region, then in an apical and submembranous position just before its secretion into the perivitelline space. The protein is synthesized in a sharply delimited continuous territory spanning about 70% of the blastula. Comparison of the size and orientation of the labelled territory in the late blastula with the fate map of the blastula stage embryo shows that the domain in which the BP10 gene is expressed corresponds to the presumptive ectoderm. Developing embryos treated with purified antibodies against the BP10 protein and with synthetic peptides derived from the EGF-like domain displayed perturbations in morphogenesis and were radialized to various degrees. These results are consistent with a role for BP10 in the differentiation of ectodermal lineages and subsequent patterning of the embryo. On the basis of these results, we speculate that the role of BP10 in the sea urchin embryo might be similar to that of tolloid in Drosophila. We discuss the idea that the processes of spatial regulation of gene expression along the animal-vegetal in sea urchin and dorsal-ventral axes in Drosophila might have some similarities and might use common elements.

Amino Acid Sequence↗
PubMed1991

A Brassica S-locus gene promoter targets toxic gene expression and cell death to the pistil and pollen of transgenic Nicotiana.

The S-locus glycoprotein gene of Brassica is derived from the genetic locus that controls the self-incompatibility response and the specific recognition between pollen and stigma. The promoter of this gene was used to direct expression of the diphtheria toxin A chain gene and the Escherichia coli beta-glucuronidase gene in transgenic Nicotiana tabacum. Expression of the promoter in cells of the pistil and in pollen suggests that a single gene may direct the self-incompatibility response in the two interacting cell types. Additionally, the fusion genes were expressed gametophytically in the heterologous host species, Nicotiana, rather than sporophytically as expected for Brassica. Thus, although the genes involved in self-incompatibility in Brassica and Nicotiana are not homologous in their coding regions, signals for expression of these genes are apparently conserved between the two genera. Our analysis of toxic gene fusion transformants shows that genetic ablation is useful for probing developmental processes and for studying temporal and spatial patterns of gene expression in plants.

Brassica↗