Universal primers for the amplification and sequence analysis of actin-1 from diverse mosquito species.

Journal of the American Mosquito Control Association
Molly StaleyB J Blitvich

Abstract

We report the development of universal primers for the reverse-transcription polymerase chain reaction (RT-PCR) amplification and nucleotide sequence analysis of actin cDNAs from taxonomically diverse mosquito species. Primers specific to conserved regions of the invertebrate actin-1 gene were designed after actin cDNA sequences of Anopheles gambiae, Bombyx mori, Drosophila melanogaster, and Caenorhabditis elegans. The efficacy of these primers was determined by RT-PCR with the use of total RNA from mosquitoes belonging to 30 species and 8 genera (Aedes, Anopheles, Culex, Deinocerites, Mansonia, Psorophora, Toxorhynchites, and Wyeomyia). The RT-PCR products were sequenced, and sequence data were used to design additional primers. One primer pair, denoted as Act-2F (5'-ATGGTCGGYATGGGNCAGAAGGACTC-3') and Act-8R (5'-GATTCCATACCCAGGAAGGADGG-3'), successfully amplified an RT-PCR product of the expected size (683-nt) in all mosquito spp. tested. We propose that this primer pair can be used as an internal control to test the quality of RNA from mosquitoes collected in vector surveillance studies. These primers can also be used in molecular experiments in which the detection, amplification or silencing of a ubiquitously expressed mosqu...Continue Reading

References

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Citations

Feb 23, 2012·Phytopathology·Diego F Quito-AvilaRobert R Martin
Sep 23, 2011·Journal of Medical Entomology·Rungrat SaiyasombatBradley J Blitvich
Apr 14, 2016·Memórias do Instituto Oswaldo Cruz·Rosalía Pérez-CastroHans J Overgaard
Nov 22, 2017·Biomédica : revista del Instituto Nacional de Salud·Myriam Lucía Velandia-RomeroMaría Inés Matiz
Jul 12, 2018·Tropical Medicine and Health·Hanako IwashitaNoboru Minakawa
Sep 15, 2021·Vector Borne and Zoonotic Diseases·Ashok MunivenkatappaPatil OmPrakash
Sep 17, 2021·Virology Journal·Tomo DaidojiTakaaki Nakaya

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Datasets Mentioned

BETA
U02933.1
GQ981439
GQ981468

Methods Mentioned

BETA
PCRs
PCR

Software Mentioned

MRBAYES

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