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J A Seawright

Publications and source records attributed to J A Seawright.

At least 19 recordsLinked to original sources
PubMed1998

Species-specific repeat units in the intergenic spacer of the ribosomal RNA cistron of Anopheles aquasalis Curry.

A genomic DNA library of Anopheles aquasalis Curry was screened for clones that hybridized more intensely to DNA from A. aquasalis than to DNA from A. benarrochi Gabaldon, Cova Garcia, and Lopez, A. konderi Galvao and Damasceno, A. nuneztovari Gabaldon cytotypes A, B, and C, A. oswaldoi (Peryassu), A. rangeli Gabaldon, Cova Garcia, and Lopez, or A. trinkae Faran. Two specific clones (2.5 kilobasepairs [kbp] and 3.0 kbp) from A. aquasalis were isolated. Both A. aquasalis-specific clones were from the intergenic spacer region of the ribosomal RNA (rRNA) cistron. Upon digestion with Rsa I, a 900-bp fragment from the clone AA-1 hybridized specifically to A. aquasalis DNA. Analysis of the DNA sequence of this fragment revealed four tandemly repeated 36-bp units. Three of these repeat units were identical, and the fourth was 94% identical to the others. The DNA sequence of a highly conserved region of these repeats was used to synthesize an oligonucleotide probe specific to A. aquasalis.

Animals↗
PubMed1997

Analysis of the Anopheles (Anopheles) quadrimaculatus complex of sibling species (Diptera: Culicidae) using morphological, cytological, molecular, genetic, biochemical, and ecological techniques in an integrated approach.

The Anopheles quadrimaculatus complex of 5 cryptic species (i.e., An. diluvialis Reinert, new species; An. inundatus Reinert, new species; An. maverlius Reinert, new species; An. quadrimaculatus Say; An. smaragdinus Reinert, new species) is analyzed using multiple techniques, including morphological, cytological, molecular, genetic, biochemical, and ecological procedures. All life stages (egg, 4th-instar larva, pupa, and female and male adults) are described using morphological features, and pertinent stages or structures are illustrated. A neotype for An. quadrimaculatus is designated, and the synonymy of An. annulimanus Van der Wulp is confirmed. Several new morphological features are described. New and summarized data from published literature on hybridization, cytological, electrophoretic, molecular, and cuticular hydrocarbon studies are included. Immature and adult bionomics are given. The geographic distribution for each species is listed and shown on maps. Procedures for collecting, processing, and rearing specimens are described. Keys using morphological characters are included for the eggs, 4th-instar larvae, pupae, adult females, and male genitalia. Also, a biochemical key for the 5 species is included. Color and pattern variations of larvae and pupae are discussed.

Animals↗
PubMed1997

Analysis of the cuticular hydrocarbons among species of the Anopheles quadrimaculatus complex (Diptera: Culicidae).

Cuticular hydrocarbons were extracted from females of 5 species of the Anopheles quadrimaculatus complex and studied by gas chromatography and mass spectrometry. The data were analyzed by multivariate techniques to determine the degree of divergence in hydrocarbon patterns and to develop models that allow the discrimination of these species. Anopheles quadrimaculatus Say, Anopheles smaragdinus Reinert, and Anopheles maverlius Reinert could be separated at 100% from each other and from Anopheles diluvialis Reinert and Anopheles inundatus Reinert; however, separation of An. diluvialis from An. inundatus was 80% using a 2-way model.

Animals↗
PubMed1996

Precise limitation of concerted evolution to ORFs in mosquito Hsp82 genes.

Two Hsp82 genes were isolated from the malaria vector Anopheles albimanus in a single lambda phage clone. The two genes are in a head-to-head arrangement separated by approx. 0.9 kbp. Northern hybridizations and 5' RACE demonstrate that both genes are transcribed, have moderate levels of constitutive transcription, and are also heat-inducible with maximum transcript accumulation occurring after 40 degrees C heat shocks. Both genes have typical heat-shock promoters and conserved intron boundaries in the untranslated leaders. The open reading frames are 99.6% identical differing in only nine silent nucleotide positions in the 2166 bp ORFs. However, precisely outside the ORFs, the flanking DNA of the two genes shows no evidence of common derivation. The high degree of identity between the two ORFs appears to be a result of gene conversion occurring by a process similar to that previously suspected in the A. albimanus Hsp70 genes and several D. melanogaster genes arranged as palindromes. This process probably involves a stem-loop intermediate and is restricted in extent by flanking sequence divergence. These Hsp82 genes clearly demonstrate the extreme precision with which gene conversion can lead to protein-coding-region homogeneity yet allow flanking DNA divergence.

Amino Acid Sequence↗
PubMed1995

Genetic differentiation and diagnostic loci of Anopheles nuneztovari, An. trinkae, and An. rangeli (Diptera: Culicidae).

Samples of Anopheles rangeli Gabaldon, Cova Garcia & Lopez, An. trinkae Causey, and An. nuneztovari Gabaldon from Venezuela, Ecuador, Brazil, and Bolivia were analyzed for genetic variability at 24 enzyme loci. Estimates of genetic variability for An. rangeli and An. trinkae from Ecuador and for An. nuneztovari in Venezuela had the following ranges: 46-58% polymorphic loci, 1.7-2.0 (SEM = 0.1-0.3) mean number of alleles per locus, and 0.069-0.113 (SEM = 0.03-0.04) expected mean heterozygosity. Genetic variability estimates of An. rangeli from Bolivia were 20.8-29.2% polymorphic loci, 1.2-1.6 (SEM = 0.1-0.2) mean number of alleles per locus, and 0.037-0.054 (SEM = 0.02-0.03) expected mean heterozygosity. The estimated genetic distance between An. rangeli and An. trinkae ranged from 0.149 to 0.197. The genetic distance between these 2 species and An. nuneztovari ranged from 0.319 to 0.440. Although there were allele frequency differences at some loci between samples of An. nuneztovari sampled from either side of the Andes Mountains in Venezuela, there were no diagnostic loci and the estimated genetic distance was only 0.023. Seven enzyme loci were diagnostic between An. nuneztovari and one or both of its sister species: Acon-2, Ao, Hk-1, Idh-2, Me, Pgi, and Pgm. The diagnostic loci Hk-1 and Acon-2 were not polymorphic in any species. An. rangeli and An. trinkae can be distinguished by the diagnostic loci Ao, Idh-2, and Me-1, and with a 97% probability by Pgm. Distance Wagner and unweighted pair-group method with arithmetic averaging analyses support a close phylogenetic relationship between An. trinkae and An. rangeli.

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PubMed1993

Sequence of a DNA probe specific for Anopheles quadrimaculatus species A (Diptera: Culicidae).

The nucleotide sequence was determined for a portion of a 12-kb genomic DNA clone specific for Anopheles quadrimaculatus species A. Four short, internally repeated sequences were identified. Synthetic oligonucleotide probes were prepared based on these four repeats. The oligonucleotides are highly specific and can be reliably used to separate individuals of An. quadrimaculatus species A from members of other species of the complex.

Animals↗
PubMed1993

The Hsp70 heat-shock gene family of the mosquito Anopheles albimanus.

Four Hsp70 genes of the malaria vector Anopheles albimanus were isolated from a genomic DNA library as two non-overlapping clones each containing a pair of divergently transcribed genes having 75% DNA sequence similarity to the protein-coding regions of the Drosophila melanogaster Hsp70 genes. The clones were assigned to two loci on chromosome 2R by in situ hybridization. These clones hybridize strongly to heat-shock but only weakly to non-shocked mosquito RNA. The Hsp70 gene family of A. albimanus is undergoing concerted evolution probably by gene conversion. The general arrangement of the genes suggests that divergently transcribed pairs of genes at two loci is an ancient Dipteran arrangement predating the Nematocera/Cyclorrapha divergence.

Animals↗
PubMed1993

The mitochondrial genome of Anopheles quadrimaculatus species A: complete nucleotide sequence and gene organization.

The complete sequence (15,455 bp) of the mitochondrial DNA of the mosquito Anopheles quadrimaculatus species A is reported. This genome is compact and very A+T rich (77.4% A+T). It contains genes for 2 ribosomal RNAs (rRNAs), 22 transfer RNAs (tRNAs), and 13 subunits of the mitochondrial inner membrane respiratory complexes. The gene arrangement is the same as in Drosophila yakuba, except that the positions of two contiguous tRNAs are reversed and a third tRNA is transcribed from the complementary strand. Protein-coding genes, rRNAs, and most tRNAs were similar to D. yakuba. Two tRNAs had nonstandard secondary structures comparable with those of nematode mitochondrial tRNAs. The very small putative control region (625 bp) contains no sequence motifs similar to those used in vertebrates and other insects for initiation of transcription and replication.

Amino Acid Sequence↗
PubMed1993

Multiple-technique identification of sibling species of the Anopheles quadrimaculatus complex.

In the past, most researchers used a single technique for identification of cryptic taxa, population structures, biosystematics, and phylogenetic studies. Our experience with the Anopheles quadrimaculatus complex shows the importance of using several methods on individual mosquitoes. This approach consists of analysis of the polytene chromosomes in ovarian nurse cells, gas chromatographic profiles of cuticular hydrocarbons, isozyme electrophoresis, and restriction site analysis of mitochondrial or genomic DNA. We recommend use of this multiple-technique approach when analyzing feral populations for the first time, or for correlating information obtained by investigators using different techniques.

Animals↗
PubMed1993

A new cytotype of Anopheles nuneztovari from western Venezuela and Colombia.

Cytogenetic analysis of the larval polytene chromosomes of Anopheles nuneztovari from 5 collection sites in Táchira and Zulia states northwest of the Andean Cordillera in western Venezuela and from 2 sites in the Department of Valle, western Colombia, revealed what appears to be a distinctive cytotype informally designated as An. nuneztovari C. Its chromosomes are homosequential with those of An. nuneztovari B from western Venezuela southeast of the Cordillera but differ in the presence of a well-defined chromocenter and unique inversion polymorphisms. The large complex inversion in western Venezuela, 2Lb, is present at a frequency of 0.263 and deviates significantly from Hardy-Weinberg equilibrium in 3 of the 5 sites. Two smaller inversions (2Lc and 2Ld) that are included in 2Lb are present in the Colombian samples at a frequency of 0.300.

Animals↗
PubMed1993

In situ hybridization mapping of histone genes in Anopheles albimanus.

The histone genes of Anopheles albimanus were mapped by in situ hybridization to 6 bands in Region 34A on the right arm of chromosome 3. A genomic library was made by cloning fragments of 15 to 23 kb (derived from partial EcoRI digestion) into the phage vector, EMBL4, and probed with the histone gene repeat of Drosophila melanogaster. Thirty-two phages containing histone gene sequences were isolated from about 10(5) plaque-forming units (pfu). Complete EcoRI digestion of DNA from 5 of the 32 recombinant phages and the genomic DNA of An. albimanus yielded a single 3.84-kb fragment that contained sequences homologous to the 5 histone genes of D. melanogaster. This 3.84-kb unit of mosquito histone genes was subcloned into puc19 plasmid, and the resulting clone (palbi34A) was used for in situ hybridization to salivary gland chromosomes.

Animals↗
PubMed1992

Mitochondrial and ribosomal DNA variation among members of the Anopheles quadrimaculatus (Diptera: Culicidae) species complex.

The extent of intra- and inter-specific variation in mitochondrial DNA and nuclear ribosomal RNA gene restriction sites was determined for the four sibling species of the Anopheles quadrimaculatus complex. Individual mosquitoes were identified by allozyme analysis according to previously published keys, and the total genomic DNA of these same individuals was then cleaved with restriction enzymes. Restriction maps of mitochondrial DNA, including the positions of variable sites, were constructed for each species. No evidence for interspecific hybridization was found in the populations surveyed. There was little variation in restriction patterns within any given species, but differences occurred among the four. Three restriction enzymes (AvaI, HindIII, and PvuII) yielded species-specific DNA restriction patterns for the mitochondrial DNA, while AvaI and HindIII produced diagnostic patterns for the ribosomal DNA. Thus, restriction patterns were very useful for detecting cryptic species but less appropriate than isozymes for studying genetic structure of populations within species.

Animals↗
PubMed1992

Quick blots and nonradioactive detection of DNA probes for the identification of mosquitoes.

The quick blot protocol is an improved technique for preparing crude insect homogenates for hybridization to nucleic acid probes. Individual insects are ground in wells of a microtiter plate and transferred to a dot blot manifold. This allows preparation of multiple filters and provides uniformity and an orderly arrangement of samples. The high background detection resulting from use of crude insect homogenates with nonradioactive detection systems was eliminated by incubating quick blot filters in a laundry stain remover containing proteases. We used mosquito species-specific DNA probes to demonstrate the effectiveness of nonradioactive DNA labeling systems with quick blots.

Animals↗
PubMed1992

Comparative susceptibility of species A, B and C of Anopheles quadrimaculatus complex to infection with subperiodic Brugia malayi and Brugia pahangi (Nematoda: Filarioidea).

Susceptibilities of natural populations of sibling species A, B and C of the Anopheles quadrimaculatus complex and the colonized strain A to subperiodic Brugia malayi and Brugia pahangi were compared. All 3 sibling species showed varying degrees of susceptibility to both B. pahangi and B. malayi, and they were considerably more susceptible to B. pahangi than to B. malayi. The rate and intensity of infection to B. pahangi were highest for species A (66.2% and 7.4 L3/female, respectively) and lowest for species B (21.3% and 1.7 L3/female). For B. malayi these values were higher for species A (29.7% and 1.84 L3/female) than for species B (13.3% and 0.86 L3/female) and C (12.6% and 0.75 L3/female). The colonized strain A of An. quadrimaculatus was significantly more susceptible to both Brugia species than the natural populations of sibling species A, B and C.

Animals↗
PubMed1991

Diagnostic characterization of Anopheles freeborni and An. hermsi by hybrid crosses, frequencies of polytene X chromosomes and rDNA restriction enzyme fragments.

A polytene chromosome analysis was prepared from Anopheles freeborni collected from 25 locations in north and central California, and parts of Washington and Oregon. The X chromosome banding pattern, thought previously to be specific to An. hermsi, was common in mosquitoes collected from foothill regions in California, and in all samples from Washington and Oregon. At some of these locations, many mosquitoes had heterokaryotypes for the inversion that distinguishes the X chromosome of An. freeborni from that of An. hermsi. Use of rDNA restriction site analysis, and the results from crossing of different strains bearing either type of X chromosome, showed that An. hermsi does not have a unique or diagnostic X chromosome. Anopheles hermsi was collected in San Mateo County, CA, which is now the northernmost known limit of this species. Crossing studies, or the examination of rDNA restriction enzyme profiles, are presently the only means of identifying An. hermsi.

Animals↗