A Ribosomal RNA Gene Probe Differentiates Member Species of the Anopheles gambiae Complex

Abstract

A 0.59 kilobase DNA fragment cloned from an rDNA cistron of the mosquito Anopheles gambiae can be used as a probe to differentiate between A. gambiae, A. arabiensis, and A. melas, three morphologically identical sibling species in the A. gambiae complex which otherwise can be reliably distinguished only by polytene chromosome banding patterns. Although all are important (and often sympatric) African malaria vectors, their relative roles in malaria transmission have thus far been difficult to assess. The probe, an EcoRI-SalI fragment from the 3' end of the 28S beta coding region of the cistron, is present in all three species, but the species differ uniquely with respect to the location of an EcoRI site in the nontranscribed spacer (NTS) downstream of the fragment. We have routinely used the probe to identify A. gambiae complex mosquitoes to species on the basis of genomic DNA extracted from individual air dried specimens. A single mosquito abdomen provides more than sufficient DNA for the assay, and neither eggs nor a bloodmeal in the abdomen interfere with DNA yield. Moreover, the DNA extraction procedure does not degrade the bloodmeal IgG, so the residual protein pellet can be used to identify the mosquito bloodmeal source. Since the rDNA cistron organization as detected by the probe does not differ between male and female mosquitoes, the probe can be used for either sex. Preliminary experiments show that the probe is equally useful for mosquito larvae and pupae.

Análisis de la estructura primaria y secundaria del ARN de transferencia mitocondrial para serina en siete especies de Lutzomyia

“…La extracción del ADN se desarrolló de acuerdo con el protocolo de Collins et al (22), con las siguientes modificaciones. Los segmentos corporales seleccionados de cada individuo se maceraron con micropistilos estériles en 60 µl de la solución tampón de lisis (0,08 NaCl, 0,16 M sacarosa, 0,06 M EDTA, 0,5% SDS, 0,1 M Tris-HCL a pH de 7,5), incubando a una temperatura de 65°C por 30 minutos.…”

Systematics and Population Level Analysis of Anopheles darlingi

“…Population level -DNA was extracted from individual wild-caught mosquitoes following the protocol in Collins et al (1987). These mosquitoes were collected from BZ (Belize), EJ (El Juval, Trujillo, Venezuela), AY (Puerto Ayacucho, Amazonas, Venezuela), FG (French Guiana), IQ (Iquitos, Peru), GU (Guayaramerín, Bolivia), BV (Boa Vista, Roraima, Brazil), MC (Macapá, Amapá, Brazil), CP (Capanema, Pará, Brazil), NS (Tefé, Amazonas, Brazil), IT (Itacoatiara, Amazonas, Brazil), AB (Porto Velho, Rondônia, Brazil), PX (Peixoto de Azevedo, Mato Grosso, Brazil), SP (Araraquara, São Paulo, Brazil) and DO (Dourado, São Paulo, Brazil) ( Fig.).…”

Variation in cuticular hydrocarbons among strains of the Anopheles gambiae sensu stricto by analysis of cuticular hydrocarbons using gas liquid chromatography of larvae

“…gambiae sensu stricto (s.s.) (Gale & Crampton 1987) as well as (g) using ribosomal RNA gene probe to differentiate members of the An. gambiae complex (Collins et al 1987). …”