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Biomedical subjects

B Burr

Publications and source records attributed to B Burr.

At least 19 recordsLinked to original sources

A combined RFLP-SSR-AFLP map of tetraploid cotton based on a Gossypium hirsutum x Gossypium barbadense backcross population.

An interspecific Gossypium hirsutum x Gossypium barbadense backcross population of 75 BC1 plants was evaluated for 1014 markers. The map consists of 888 loci, including 465 AFLPs, 229 SSRs, 192 RFLPs, and 2 morphological markers, ordered in 37 linkage groups that represent most if not all of the 26 chromosomes, altogether spanning 4400 cM. Loci were not evenly distributed over linkage groups, and 18 of the 26 long groups had a single dense region. This paper proposes a partially revised list of the 13 pairs of homoeologous A/D chromosomes of the 2n = 4x = 52 tetraploid cotton genome. The major revisions, which involve the c3-c17, c4-c22, c5-D08, and c10-c20 homoeologous pairs, are based on the mapping of 68 SSR and RFLP loci with a known chromosome assignment, as well as on comparative alignments with previously published G. hirsutum x G. barbadense maps. The overall congruency in the locus orders and distances of common SSR and RFLP loci in these maps allows for an estimation of the consensus length that reaches a minimum of 5500 cM, and is encouraging for future efforts aimed at developing an integrated map of cultivated cotton. The present map also provides a firm framework for precision mapping of Mendelian components of quantitative traits in cotton

Chromosome Mapping↗

Commonly used antibacterial and antifungal agents for hospitalised paediatric patients: implications for therapy with an emphasis on clinical pharmacokinetics.

Due to normal growth and development, hospitalised paediatric patients with infection require unique consideration of immune function and drug disposition. Specifically, antibacterial and antifungal pharmacokinetics are influenced by volume of distribution, drug binding and elimination, which are a reflection of changing extracellular fluid volume, quantity and quality of plasma proteins, and renal and hepatic function. However, there is a paucity of data in paediatric patients addressing these issues and many empiric treatment practices are based on adult data. The penicillins and cephalosporins continue to be a mainstay of therapy because of their broad spectrum of activity, clinical efficacy and favourable tolerability profile. These antibacterials rapidly reach peak serum concentrations and readily diffuse into body tissues. Good penetration into the cerebrospinal fluid (CSF) has made the third-generation cephalosporins the agents of choice for the treatment of bacterial meningitis. These drugs are excreted primarily by the kidney. The carbapenems are broad-spectrum beta-lactam antibacterials which can potentially replace combination regimens. Vancomycin is a glycopeptide antibacterial with gram-positive activity useful for the treatment of resistant infections, or for those patients allergic to penicillins and cephalosporins. Volume of distribution is affected by age, gender, and bodyweight. It diffuses well across serous membranes and inflamed meninges. Vancomycin is excreted by the kidneys and is not removed by dialysis. The aminoglycosides continue to serve a useful role in the treatment of gram-negative, enterococcal and mycobacterial infections. Their volume of distribution approximates extracellular space. These drugs are also excreted renally and are removed by haemodialysis. Passage across the blood-brain barrier is poor, even in the face of meningeal inflammation. Low pH found in abscess conditions impairs function. Toxicity needs to be considered. Macrolide antibacterials are frequently used in the treatment of respiratory infections. Parenteral erythromycin can cause phlebitis, which limits its use. Parenteral azithromycin is better tolerated but paediatric pharmacokinetic data are lacking. Clindamycin is frequently used when anaerobic infections are suspected. Good oral absorption makes it a good choice for step-down therapy in intra-abdominal and skeletal infections. The use of quinolones in paediatrics has been restricted and most information available is in cystic fibrosis patients. High oral bioavailability is also important for step-down therapy. Amphotericin B has been the cornerstone of antifungal treatment in hospitalised patients. Its metabolism is poorly understood. The half-life increases with time and can be as long as 15 days after prolonged therapy. Oral absorption is poor. The azole antifungals are being used increasingly. Fluconazole is well tolerated, with high bioavailability and good penetration into the CSF. Itraconazole has greater activity against aspergillus, blastomycosis, histoplasmosis and sporotrichosis, although it's pharmacological and toxicity profiles are not as favourable.

Anti-Bacterial Agents↗

International Rice Genome Sequencing Project: the effort to completely sequence the rice genome.

The International Rice Genome Sequencing Project (IRGSP) involves researchers from ten countries who are working to completely and accurately sequence the rice genome within a short period. Sequencing uses a map-based clone-by-clone shotgun strategy; shared bacterial artificial chromosome/P1-derived artificial chromosome libraries have been constructed from Oryza sativa ssp. japonica variety 'Nipponbare'. End-sequencing, fingerprinting and marker-aided PCR screening are being used to make sequence-ready contigs. Annotated sequences are immediately released for public use and are made available with supplemental information at each IRGSP member's website. The IRGSP works to promote the development of rice and cereal genomics in addition to producing genome sequence data.

Chromosome Mapping↗

Simple inheritance of key traits distinguishing maize and teosinte.

The segregation of key traits distinguishing maize and teosinte was analyzed in three F2 and three backcross populations derived from crosses of the modern maize inbred T232 with Zea mays ssp. parviglumis. These traits were (i) paired vs. single female spikelets; (ii) two-ranked vs. many-ranked ears; (iii) non-indurated vs. indurated glumes; (iv) inclination of the kernels toward the rachis, and (v) distichous vs. polystichous central staminate spike. All traits showed a simple mode of inheritance except for paired female spikes, which appeared to be controlled by two genes. The loci controlling these major changes were mapped with RFLP markers to four chromosomal regions. These results support the suggestion that maize became differentiated from teosinte with as few as five major gene changes.

Chromosome Mapping↗

Cloning and characterization of a maize cDNA encoding phytoene desaturase, an enzyme of the carotenoid biosynthetic pathway.

To study regulation of the plastid-localized maize carotenoid biosynthetic pathway, a cDNA encoding phytoene desaturase (PDS) was isolated and characterized. The DNA sequence of the maize Pds cDNA was determined and compared with available dicot Pds genes. The deduced PDS protein, estimated at 64.1 kDa (unprocessed), had a dinucleotide binding domain and conserved regions characteristic of other carotene desaturases. Alignment of available PDS sequences from distantly related organisms suggests that Pds has potential as a phylogenetic tool. By use of heterologous complementation in Escherichia coli, maize PDS was shown to catalyze two desaturation steps converting phytoene to zeta-carotene. RFLP (restriction fragment length polymorphism) mapping was used to place Pds on chromosome 1S near viviparous5 (vp5), and RT-PCR (reverse-transcriptase polymerase chain reaction) analysis indicated reduced Pds transcript in vp5 mutant relative to normal endosperm. Other phytoene-accumulating mutant endosperms, vp2 and white3 (w3), showed no difference in Pds transcript accumulation as compared with normal endosperm counterparts. RT-PCR analysis of Pds transcript accumulation in developing endosperm showed Pds was constitutively expressed. Therefore, endosperm carotenogensis is not regulated by increasing the level of Pds transcripts.

Amino Acid Sequence↗

The maize repressor-like gene intensifier1 shares homology with the r1/b1 multigene family of transcription factors and exhibits missplicing.

The recessive mutation intensifier1 of maize apparently causes an overall increase in flavonoid production in the aleurone. The mechanism by which this is achieved is not understood. We have succeeded in cloning the intensifier1 gene by transposon tagging with Suppressor-mutator and found, by sequence analyses, that it shares homology with known transcription factors in the anthocyanin pathway, in particular the r1/b1 multigene family in maize. Two cDNAs and a genomic clone were completely sequenced, and together they showed that the transcripts were misspliced. The frequency of missplicing was investigated by polymerase chain reaction analyses and sequencing of the individual introns. These studies indicate that very little functional transcript was made. Indeed, missplicing may be a mechanism for reducing the levels of a transcription factor that, when present, acts as a repressor of anthocyanin biosynthesis.

Amino Acid Sequence↗

Maize anthocyanin regulatory gene pl is a duplicate of c1 that functions in the plant.

Genetic studies in maize have identified several regulatory genes that control the tissue-specific synthesis of purple anthocyanin pigments in the plant. c1 regulates pigmentation in the aleurone layer of the kernel, whereas pigmentation in the vegetative and floral tissues of the plant body depends on pl. c1 encodes a protein with the structural features of eukaryotic transcription factors and functions to control the accumulation of transcripts for the anthocyanin biosynthetic genes. Previous genetic and molecular observations have prompted the hypothesis that c1 and pl are functionally duplicate, in that they control the same set of anthocyanin structural genes but in distinct parts of the plant. Here, we show that this proposed functional similarity is reflected by DNA sequence homology between c1 and pl. Using a c1 DNA fragment as a hybridization probe, genomic and cDNA clones for pl were isolated. Comparison of pl and c1 cDNA sequences revealed that the genes encode proteins with 90% or more amino acid identity in the amino- and carboxyl-terminal domains that are known to be important for the regulatory function of the C1 protein. Consistent with the idea that the pl gene product also acts as a transcriptional activator is our finding that a functional pl allele is required for the transcription of at least three structural genes in the anthocyanin biosynthetic pathway.

Amino Acid Sequence↗

Changes in state of the Wx-m5 allele of maize are due to intragenic transposition of Ds.

The molecular basis for the unusual phenotype conditioned by the waxy(Wx)-m5 Ds allele has been elucidated. Unlike most Ds alleles, Wx-m5 is phenotypically wild-type in the absence of Ac. We find that the Wx-m5 gene contains a 2-kb Ds element at -470 relative to the start of Wx transcription, representing the most 5' insertion of any transposable element allele characterized to date in plants. Despite its wild type phenotype, Wx-m5 has reduced levels of Wx enzymatic activity indicating that Ds insertion influences Wx gene expression. In the presence of Ac, Wx-m5 kernels have sectors of null expression on a wild-type background and give rise to stable wx and unstable wx-m germinal derivatives. Seventeen of 20 derivatives examined are wx-m alleles and at least 15 of these appear to result from intragenic transposition of Ds from -470 to new sites within the Wx gene. Three wx-m alleles contain two Ds elements, one at -470 and a second in Wx coding sequences. Surprisingly, only 3 out of 20 derivatives are stable wx mutants and these have sustained gross rearrangements of Wx and flanking sequences. For most other maize transposable element alleles somatic sectors and germinal derivatives usually arise following element excision or deletions of element sequences. In contrast, element insertion following intragenic transposition is apparently responsible for most of the somatic sectors and germinal derivatives of Wx-m5.

Alleles↗

Pinning down loose ends: mapping telomeres and factors affecting their length.

A degenerately repeated sequence, proximal to the telomere heptanucleotide repeat in maize, contains restriction enzyme sites that permit the separation of telomeres from the rest of the chromosomes. Probing with a telomere-specific oligonucleotide revealed genotype-dependent telomere lengths that vary more than 25-fold in maize among the 22 inbreds that have been surveyed. These lengths were found to segregate reproducibly in a recombinant inbred family where 50% of the variation can be accounted for by three loci. The dynamic control over telomere length in maize appears to act rapidly to achieve new genotypically determined telomere lengths in the F1. Clones of telomere proximal sequences were used to map restriction fragment length loci at the distal ends of eight of 20 chromosome arms.

Base Sequence↗

Recombinant inbreds for molecular mapping in maize: theoretical and practical considerations.

Molecular mapping is rapidly being extended to more species as one result of the current emphasis on genome initiatives. The choice of the population used for mapping can have important consequences on the efficiency and accessibility of the mapping information. Recombinant inbred lines offer certain advantages over other mapping populations for many species. These more or less permanent populations permit many geneticists to contribute to the mapping effort and to profit from each other's work. Recombinant inbred lines are used extensively in mouse genetics and have been used in maize to compile a detailed molecular map.

Chromosome Mapping↗

An arginine to lysine substitution in the bZIP domain of an opaque-2 mutant in maize abolishes specific DNA binding.

The opaque-2 (o2) locus in maize encodes a transcription factor involved in the regulation of zein storage proteins. We have shown previously that the O2 protein contains a leucine zipper domain that binds to promoters of 22-kD zein genes. In this paper we characterize an EMS-induced o2 allele, o2-676, that causes a 50% reduction in zein. We have found that the o2-676 mutant protein does not show specific recognition of zein promoter fragments because of the substitution of a lysine residue for an arginine residue within the bZIP domain of o2-676. This particular arginine is conserved within the bZIP domains of all mammalian, fungal, and plant DNA binding proteins of this class. The correlation between this mutation in o2 and the altered pattern of zein expression strongly suggests that O2 regulates transcription of certain members of the zein multigene family through direct interaction with the zein promoters and not through the transcriptional activation of some other regulator of zein gene expression.

Amino Acid Sequence↗

Maize regulatory gene opaque-2 encodes a protein with a "leucine-zipper" motif that binds to zein DNA.

The opaque-2 locus (o2) in maize regulates the expression of many members of the zein multigene family of storage proteins. cDNA clones for a wild-type allele of the (o2) locus (O2) were isolated from a maize endosperm cDNA library and sequenced. We found a 258-nucleotide 5' leader sequence containing three short open reading frames followed by a sequence specifying a protein of 437 amino acids. The presumptive amino acid sequence of the protein (O2) specified by the O2 cDNA contains a "leucine-zipper" domain characteristic of some mammalian and fungal transcription activation factors. lacZ-O2 fusion constructs, using nearly the entire coding region of O2 or only a fragment specifying the leucine-zipper domain, were expressed in Escherichia coli. In an in vitro binding assay, the beta-galactosidase-O2 fusion proteins bound to two specific regions on the 5' side of the coding sequence in a zein genomic clone. This suggests that the O2 protein affects zein transcription through direct interaction with one or more zein promoter elements.

Amino Acid Sequence↗

Advantages and limitations of using Spm as a transposon tag.

Transposon tagging has become the method of choice for isolating genes whose products are in low abundance. We have recently used the transposable element Spm to tag and clone maize regulatory loci. Our choice of Spm was dictated by several factors: The frequency of transposition of Spm is high enough to obtain detectable transposition events, into loci affecting kernel traits, in populations of less than 10(6) seed. Although the copy number of Spm is high in the maize genome, insertions into the gene of interest can be distinguished from other Spm copies by digesting DNAs from segregating populations with methyl-sensitive restriction enzymes, and hybridizing with Spm-specific probes. Since all members of the Spm family thus far examined share DNA homology, hybridization with appropriate probes allows detection of insertions of both autonomous and defective elements. Thus, if a mutable allele can be shown to be under Spm control, one can be reasonably confident of successfully cloning that allele.

Cloning, Molecular↗

Activation of silent transposable elements.

It is well known among maize geneticists that agents that cause chromosome breakage can activate quiescent transposable elements. However, other than temporarily relieving position effect, it is difficult to understand how these events can lead directly to activation. One possibility is that chromosome breakage can initiate a process in the cell resulting in a higher rate of spontaneous mutation. Such a system could be analogous to the SOS response of Escherichia coli in which an error-prone repair system is induced. Chemical mutagens that cause little chromosome breakage but add bulky adducts to the DNA can induce the SOS response. In seed homozygous for a1-m2(8004), wx-m8, no active Spm, that had been treated with ethyl methanesulfonate, we observed activation of Spm at the rate of 1.1 x 10(-4). The spontaneous rate of activation in this material was 1.2 x 10(-5). Most of the activation events occurred as single kernels. This result contrasts with sectors covering at least one-eighth of the ear that would have been expected if activation had occurred as a direct result of mutagenesis in the mature kernel. The late timing of these events suggests that the activation, in most instances, may not be the direct result of chemical mutagenesis.

Crosses, Genetic↗

Gene mapping with recombinant inbreds in maize.

Recombinant inbred lines of maize have been developed for the rapid mapping of molecular probes to chromosomal location. Two recombinant inbred families have been constructed from F2 populations of T232 X CM37 and CO159 X Tx303. A genetic map based largely on isozymes and restriction fragment length polymorphisms has been produced that covers virtually the entire maize genome. In order to map a new gene, an investigator has only to determine its allelic distribution among the recombinant inbred lines and then compare it by computer with the distributions of all previously mapped loci. The availability of the recombinant inbreds and the associated data base constitute an efficient means of mapping new molecular markers in maize.

Chromosome Mapping↗

Transposon tagging and molecular analysis of the maize regulatory locus opaque-2.

Genetic analyses suggested that the opaque-2 (o2) locus in maize acts as a positive, transacting, transcriptional activator of the zein seed storage-protein genes. Because isolation of the gene is requisite to understanding the molecular details of this regulation, transposon mutagenesis with the transposable element suppressor-mutator (Spm) was carried out, and three mutable o2 alleles were obtained. One of these alleles contained an 8.3-kilobase autonomous Spm, another a 6.8-kilobase nonautonomous Spm, and the third an unidentified transposon that is unrelated to Spm. A DNA sequence flanking the autonomous Spm insertion was verified to be o2-specific and provided a probe to clone a wild-type allele. Northern blots indicated that the gene is expressed in wild-type endosperm but not in leaf tissues or in endosperms homozygous for a mutant allele of the O2 gene. A transcript was detected in endosperms homozygous for mutations at opaque-7 and floury-2, an indication that O2 expression is independent of these two other putative regulators of zein synthesis.

Alleles↗

Molecular analysis of the maize anthocyanin regulatory locus C1.

The C1 gene of maize plays a regulatory role in the production of anthocyanin pigments in the aleurone layer of the endosperm. As an initial step toward understanding the molecular details of how C1 controls pigment biosynthesis, we cloned the C1 gene. This was accomplished by first cloning a mutable allele of C1, c1-m5, which contains the transposable element Spm. A combination of molecular and genetic analysis was used to identify the Spm at the C1 locus. Individual genomic DNAs from a population in which the c1-mutable phenotype was segregating with the recessive c1 phenotype were digested with methyl-sensitive restriction enzymes and probed with a small DNA fragment derived from a defective Spm. One Sal I restriction fragment complementary to the Spm probe was shown to be present in the DNA of individuals with the c1-m5 phenotype but absent from DNA of individuals with a recessive c1 phenotype. Subsequent cloning and restriction analysis of this fragment revealed sequences flanking the Spm that proved to be C1-specific. A DNA fragment derived from the flanking sequences was then used as a probe to clone the wild-type C1 gene and several additional alleles of C1, including one stable recessive, two mutations caused by Ds insertions, one mutation induced by insertion of a defective Spm, and two dominant mutations, C1-S and C1-I. RNA blot hybridization analysis of three C1 alleles indicates that C1 regulation of the Bz1 and A1 structural genes in the anthocyanin biosynthetic pathway is at the transcriptional level.

Alleles↗