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F H Collins

Publications and source records attributed to F H Collins.

At least 55 records · Page 3Linked to original sources
PubMed1997

Progress in the map-based cloning of the Anopheles gambiae genes responsible for the encapsulation of malarial parasites.

A genetically selected strain of the mosquito Anopheles gambiae, the major vector of malaria in sub-Saharan Africa, is able to encapsulate and kill Plasmodium ookinetes after they have penetrated the midgut cells and come to rest between the midgut epithelial cells and the surrounding basal lamina. The genetic basis of this phenotype has now been examined by high-resolution mapping using microsatellite loci. Results of this mapping indicate that three genes contribute to this phenotype, with one gene on the left arm of chromosome 2 accounting for the most of the effect. These genes, called Pen1, Pen2, and Pen3 (for Plasmodium encapsulation genes 1, 2 and 3) have also been physically localized to relatively small and well defined regions of the polytene chromosome complement. Strategies for cloning these genes by genetic and physical mapping methods are discussed.

Africa South of the Sahara↗
PubMed1997

Mapping a quantitative trait locus involved in melanotic encapsulation of foreign bodies in the malaria vector, Anopheles gambiae.

A Plasmodium-refractory strain of Anopheles gambiae melanotically encapsulates many species of Plasmodium, whereas wild-type mosquitoes are usually susceptible. This encapsulation trait can also be observed by studying the response of refractory and susceptible strains to intrathoracically injected CM-Sephadex beads. We report the results of broad-scale quantitative trait locus (QTL) mapping of the encapsulation trait using the bead model system. Interval mapping using the method of maximum likelihood identified one major QTL, Pen1. The 13.7-cM interval containing Pen1 was defined by marker AGH157 at 8E and AGH46 at 7A on 2R. Pen1 was associated with a maximum LOD score of 9.0 and accounted for 44% of the phenotypic variance in the distribution of phenotypes in the backcross. To test if this QTL is important for encapsulation of Plasmodium berghei, F2 progeny were infected with P. berghei and evaluated for degree of parasite encapsulation. For each of the two markers that define the interval containing Pen1, a significant difference of encapsulation was seen in progeny with at least one refractory allele in contrast with homozygous susceptible progeny. These results suggest that Pen1 is important for melanotic encapsulation of Plasmodium as well as beads.

Animals↗
PubMed1997

Patterns of mitochondrial variation within and between African malaria vectors, Anopheles gambiae and An. arabiensis, suggest extensive gene flow.

Anopheles gambiae and An. arabiensis are mosquito species responsible for most malaria transmission in sub-Saharan Africa. They are also closely related sibling species that share chromosomal and molecular polymorphisms as a consequence of incomplete lineage sorting or introgressive hybridization. To help resolve these processes, this study examined the partitioning of mtDNA sequence variation within and between species across Africa, from both population genetic and phylogeographic perspectives. Based on partial gene sequences from the cytochrome b, ND1 and ND5 genes, haplotype diversity was high but sequences were very closely related. Within species, little or no population subdivision was detected, and there was no evidence for isolation by distance. Between species, there were no fixed nucleotide differences, a high proportion of shared polymorphisms, and eight haplotypes in common over distances as great as 6000 km. Only one of 16 shared polymorphisms led to an amino acid difference, and there was no compelling evidence for nonneutral variation. Parsimony networks constructed of haplotypes from both species revealed no correspondence of haplotype with either geography or taxonomy. This trend of low intraspecific genetic divergence is consistent with evidence from allozyme and microsatellite data and is interpreted in terms of both extensive gene flow and recent range expansion from relatively large, stable populations. We argue that retention of ancestral polymorphisms is a plausible but insufficient explanation for low interspecific genetic divergence, and that extensive hybridization is a contributing factor.

Animals↗
PubMed1997

Ribosomal DNA-polymerase chain reaction assay discriminates between Anopheles quadriannulatus and An. merus (Diptera: Culicidae).

A ribosomal DNA polymerase chain reaction technique (rDNA-PCR) that distinguishes the 5 more common and widespread members of the Anopheles gambiae complex failed to consistently identify specimens of Anopheles merus Dönitz collected in South Africa and Tanzania. When the original rDNA-PCR assay was applied to field-collected specimens or specimens from laboratory colonies established from these populations, bands diagnostic of both An. merus and An. quadriannulatus (Theobald) were amplified from all individual specimens. However, all the specimens tested had the polytene chromosome banding morphology or the superoxide dismutase isozyme that were diagnostic for An. merus. Replacement of the original An. quadriannulatus-specific primer with a new primer derived from another region of the rDNA intergenic spacer resulted in an alternative rDNA-PCR assay that accurately and consistently differentiated among specimens of An. merus, An. quadriannulatus, and An. arabiensis Patton. Anopheles gambiae Giles also may be distinguished by this assay if high percentage agarose gels or gels of other matrices with better resolving powers are used.

Animals↗
PubMed1996

A shared genetic mechanism for melanotic encapsulation of CM-Sephadex beads and a malaria parasite, Plasmodium cynomolgi B, in the mosquito, Anopheles gambiae.

A Plasmodium-refractory strain of Anopheles gambiae that melanizes ookinetes and intrathoracically inoculated CM-Sephadex beads was mated to a Plasmodium-susceptible strain that does not melanize the parasite or the beads. The F1 progeny were then backcrossed to the susceptible strain. Backcross progeny were given a blood meal containing infective Plasmodium cynomolgi B, and the parasites were allowed to develop for 6-7 days, at which time the infected mosquitoes were injected with CM-Sephadex beads. The next day the mosquitoes were dissected and the beads were scored for degree of melanization while the parasites were scored for degree of encapsulation. A Spearman rank order correlation test of the degree of correlation between the bead melanization phenotype and the parasite encapsulation phenotype gave a correlation coefficient of 0.74 (P < 0.01). This strong correlation between the two melanization responses suggests that the mechanisms for differential bead and parasite melanization of these two mosquito strains share at least one major gene.

Animals↗
PubMed1996

Phylogeny of nearctic members of the Anopheles maculipennis species group derived from the D2 variable region of 28S ribosomal RNA.

Phylogenetic affinities among taxa associated with the Nearctic component of the Anopheles maculipennis species group (subgenus Anopheles) were inferred from sequence divergence in the D2 variable region of 28S ribosomal RNA. The base composition of this region had a marked GC bias which ranged from 59.9% in Anopheles walkeri to 65.1% in Anopheles punctipennis E. Although over two-thirds of the base positions in the D2 region were double-stranded (stem), substitution frequencies at single-stranded (loop) positions (0.068 over all taxa) were 2.7 times greater than at stem positions (0.025). Most mutations were point mutations and were most frequent at loop positions. In the shortest trees generated by both parsimony and distance methods, the four American species traditionally identified with the maculipennis complex (Anopheles aztecus, Anopheles earlei, Anopheles freeborni, and Anopheles occidentalis) were monophyletic with A. punctipennis E and W as sister taxa. The latter two correspond to genetically distinct forms from the eastern United States and California, respectively. The sibling species of the Anopheles quadrimaculatus complex formed a distinct clade, and A. quadrimaculatus D, with six autapomorphies, was the most divergent of these taxa. Sequence divergence between A. walkeri and the other taxa included in the study was of such magnitude as to suggest only a distant affinity to these species.

Animals↗
PubMed1996

Quetzal: a transposon of the Tc1 family in the mosquito Anopheles albimanus.

A member of the Tc1 family of transposable elements has been identified in the Central and South American mosquito Anopheles albimanus. The full-length Quetzal element is 1680 base pairs (bp) in length, possesses 236 bp inverted terminal repeats (ITRs), and has a single open reading frame (ORF) with the potential of encoding a 341-amino-acid (aa) protein that is similar to the transposases of other members of the Tc1 family, particularly elements described from three different Drosophila species. The approximately 10-12 copies per genome of Quetzal are found in the euchromatin of all three chromosomes of A. albimanus. One full-length clone, Que27, appears capable of encoding a complete transposase and may represent a functional copy of this element.

Amino Acid Sequence↗
PubMed1996

The Tryptophan oxygenase gene of Anopheles gambiae.

The Anopheles gambiae gene encoding tryptophan oxygenase, a homolog of the Drosophila melanogaster vermilion gene, has been molecularly cloned and characterized. Unlike Drosophila, where it is X-linked, the A. gambiae gene maps to chromosome 2R, subdivision 12E, by in situ hybridization to the polytene chromosomes. Of the six introns present, four are positioned identically to those of the Drosophila homolog, one is similarly positioned, and one is novel. A 1 955 nt cDNA potentially encodes a 392 amino acid protein of an estimated 45 kDa. Amino acid comparisons between the deduced protein and previously known tryptophan oxygenases revealed 74% identity between Anopheles and Drosophila, and 53% identity between Anopheles and nematode or mammalian proteins. Northern analysis detected a developmentally regulated transcript about 2 kb in length. Since this gene is known to control adult eye color in other flies, its cloning from A. gambiae provides the basis for a dominant phenotypic marker for germline transformation, one whose expression, unlike that of white, is not cell autonomous.

Amino Acid Sequence↗
PubMed1996

Genetic differentiation of Anopheles gambiae populations from East and west Africa: comparison of microsatellite and allozyme loci.

Genetic variation of Anopheles gambiae was analysed to assess interpopulation divergence over a 6000 km distance using short tandem repeat (microsatellite) loci and allozyme loci. Differentiation of populations from Kenya and Senegal measured by allele length variation at five microsatellite loci was compared with estimates calculated from published data on six allozyme loci (Miles, 1978). The average Wright's FST of microsatellite loci (0.016) was lower than that of allozymes (0.036). Slatkin's RST values for microsatellite loci were generally higher than their FST values, but the average RST value was virtually identical (0.036) to the average allozyme FST. These low estimates of differentiation correspond to an effective migration index (Nm) larger than 3, suggesting that gene flow across the continent is only weakly restricted. Polymorphism of microsatellite loci was significantly higher than that of allozymes, probably because the former experience considerably higher mutation rates. That microsatellite loci did not measure greater interpopulation divergence than allozyme loci suggested constraints on microsatellite evolution. Alternatively, extensive mosquito dispersal, aided by human transportation during the last century, better explains the low differentiation and the similarity of estimates derived from both types of genetic markers.

Alleles↗
PubMed1996

An integrated genetic map of the African human malaria vector mosquito, Anopheles gambiae.

We present a genetic map based on microsatellite polymorphisms for the African human malaria vector, Anopheles gambiae. Polymorphisms in laboratory strains were detected for 89% of the tested microsatellite markers. Genotyping was performed for individual mosquitos from 13 backcross families that included 679 progeny. Three linkage groups were identified, corresponding to the three chromosomes. We added 22 new markers to the existing X chromosome map, for a total of 46 microsatellite markers spanning a distance of 48.9 cM. The second chromosome has 57 and the third 28 microsatellite markers spanning a distance of 72.4 and 93.7 cM, respectively. The overall average distance between markers is 1.6 cM (or 1.1, 1.2, and 3.2 cM for the X, second, and third chromosomes, respectively). In addition to the 131 microsatellite markers, the current map also includes a biochemical selectable markers, Dieldrin resistance (Dl), on the second chromosome and five visible markers, pink-eye (p) and white (w) on the X, collarless (c) and lunate (lu) on the second, and red-eye (r) on the third. The cytogenetic locations on the nurse cell polytene chromosomes have been determined for 47 markers, making this map an integrated tool for cytogenetic, genetic, and molecular analysis.

Animals↗
PubMed1996

An evaluation of evolutionary constraints on microsatellite loci using null alleles.

A test to evaluate constraints on the evolution of single microsatellite loci is described. The test assumes that microsatellite alleles that share the same flanking sequence constitute a series of alleles with a common descent that is distinct from alleles with a mutation in the flanking sequence. Thus two or more different series of alleles at a given locus represent the outcomes of different evolutionary processes. The higher rate of mutations within the repeat region (10(-3) or 10(-4)) compared with that of insertion/deletion or point mutations in adjacent flanking regions (10(-9)) or with that of recombination between the repeat and the point mutation (10(-6) for sequences 100 bp long) provides the rationale for this assumption. Using a two-phase, stepwise mutation model we simulated the evolution of a number of independent series of alleles and constructed the distributions of two similarity indices between pairs of these allele series. Applying this approach to empirical data from locus AG2H46 of Anopheles gambiae resulted in a significant excess of similarity between the main and the null series, indicating that constraints affect allele distribution in this locus. Practical considerations of the test are discussed.

Alleles↗
PubMed1996

Polymerase chain reaction species diagnostic assay for Anopheles quadrimaculatus cryptic species (Diptera: Culicidae) based on ribosomal DNA ITS2 sequences.

Species-specific differences in the nucleotide sequences of the 2nd internal transcribed spacer (ITS2) of nuclear ribosomal DNA (rDNA) were used to develop a diagnostic assay based on the polymerase chain reaction (PCR) that can distinguish 4 of the 5 cryptic sibling species in the common malaria mosquito, Anopheles quadrimaculatus Say, complex. The assay requires only a small amount of tissue from an individual mosquito and a mixture of 5 PCR primers. The plus strand universal primer is derived from a sequence in the 5.8S coding region that is identical in all members of the complex. The 4 minus strand primers were selected from species-unique sequences within the ITS2 region. PCR amplification produces a different sized fragment for each of the 4 species which can be visualized readily under ultraviolet light after electrophoresis through an ethidium bromide-containing agarose gel. The assay has been developed and tested only with An. quadrimaculatus complex specimens from Florida populations.

Animals↗
PubMed1996

Validation of a ribosomal DNA-polymerase chain reaction species diagnostic assay for the common malaria mosquito (Diptera:Culicidae) sibling species complex.

A polymerase chain reaction method for identifying individuals in the Anopheles quadrimaculatus Say sibling species complex was validated for wild mosquitoes from Louisiana and Mississippi. This method distinguished An. quadrimaculatus species A, B, C, and D by detecting species-specific differences in the 2nd internal transcribed spacer of ribosomal DNA and was 100% specific and 95% sensitive.

Animals↗
PubMed1996

A review of the use of ribosomal DNA (rDNA) to differentiate among cryptic Anopheles species.

Cryptic species complexes are groups of closely related species that are difficult or impossible to distinguish by morphological traits. These complexes are known from a wide variety of arthropods and are common among the well-studied, medically-important insects. For example, many of the anopheline vectors of malaria parasites are members of cryptic species complexes. Complexes typically include both vector and non-vector species, and two or more member species are often found sympatrically. Until the late 1950, only two such Anopheles complexes were known, the A. gambiae complex from Africa and the A. maculipennis complex from Europe. Today, dozens of Anopheles cryptic species complexes are recognized, and accumulating evidence suggests that most important malaria vectors are likely to be members of such complexes. A variety of methods have been developed for identifying the species of individual specimens from these complexes, although until recently only those based on species-specific allozymes and polytene chromosome inversions were widely used. The limitations inherent in these methods have been circumvented with DNA-based procedures, which are especially useful because both sexes and all developmental stages can be identified, and DNA can be recovered from samples stored by a wide variety of simple methods. Several DNA-based identification techniques have been developed, including hybridization assays based on species-specific repeat sequences, and diagnostic PCR fragments produced either by the use of random PCR primers or by amplifying DNA with primers based on known species-specific sequences. In this review we discuss the relative marks of different methods of cryptic species identification, with emphasis on the use of ribosomal DNA as a target for species-diagnostic PCR assays.

Animals↗
PubMed1996

The polyubiquitin gene of the mosquito Anopheles gambiae: structure and expression.

The polyubiquitin gene from the mosquito Anopheles gambiae has been cloned and sequenced, and its structure is reported along with sequence analysis results. The gene consists of approximately seven tandem head-to-tail repeat units of the seventy-six amino acid-coding ubiquitin monomer. It is expressed constitutively in larvae, pupae and adults of An. gambiae, as well as in a cell line derived from this mosquito species. A probe made from a DNA fragment containing the coding region of the gene recognizes transcripts of approximately 3.6 kb and 4.4 kb in RNA isolated from all mosquito developmental stages and a unique transcript of approximately 3.0 kb in RNA from the cell line. Single monomeric units of the An. gambiae polyubiquitin gene shared from 75.9% to 85.5% identity at the DNA level with homologous sequences from other organisms ranging from yeast to man. A comparison of individual repeat units of the An. gambiae gene revealed that, in general, the 5' ends of the individual monomers are more highly conserved than the 3' ends. The gene mapped by in situ hybridization on ovarian nurse cell polytene chromosomes to a primary site at division 12C on chromosome 2R and to a secondary site at division 9C on the same chromosome.

Amino Acid Sequence↗
PubMed1995

The white gene of Ceratitis capitata: a phenotypic marker for germline transformation.

Reliable germline transformation is required for molecular studies and ultimately for genetic control of economically important insects, such as the Mediterranean fruit fly (medfly) Ceratitis capitata. A prerequisite for the establishment and maintenance of transformant lines is selectable or phenotypically dominant markers. To this end, a complementary DNA clone derived from the medfly white gene was isolated, which showed substantial similarity to white genes in Drosophila melanogaster and other Diptera. It is correlated with a spontaneous mutation causing white eyes in the medfly and can be used to restore partial eye color in transgenic Drosophila carrying a null mutation in the endogenous white gene.

ATP-Binding Cassette Transporters↗
PubMed1995

A new dominant selectable marker for genetic transformation; Hsp70-opd.

New P element plasmids containing the organophosphate-degrading gene opd as a dominant selectable marker were tested as transformation vectors in Drosophila melanogaster. One of these vectors was modified by the addition of the D. melanogaster mini-white gene as a comarker. When transformed individuals were identified using paraoxon selection for opd alone, results were similar to those obtained with mini-white. No false positives were recovered, however one strain contained the mini-white gene but inadequate resistance to survive our screening regimen due to a defective Hsp70-opd gene. Results suggest that Hsp70-opd is similar to mini-white for distinguishing transformed individuals but does not require time-consuming individual examination. Due to the mode of action of organophosphorus nerve agents, Hsp70-opd has potential as a selectable marker in numerous animals beside fruit flies.

Animals↗