Heterologous Expression of AaNa(v)1-1 in Xenopus Oocytes

AaNa_v1-1 and mutants were expressed in the Xenopus oocytes, Ovaries from oocyte-positive female Xenopus laevis purchased from Xenopus 1 (Dexter, MI). The procedures for oocyte preparation, cRNA synthesis and injection were identical to those described previously [35 (link)]. cRNA was prepared by in vitro transcription with T7 polymerase using the mMESSAGE mMACHINE high yield capped RNA kit (Ambion, Austin, TX). To enhance expression of AaNa_v1-1 and mutant channels, their cRNAs were co-injected into oocytes with Ae. aegypti tipE cRNA in the 1:1 molar ratio [36 (link), 37 (link)].

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Chen M., Du Y., Wu S., Nomura Y., Zhu G., Zhorov B.S, & Dong K. (2019). Molecular evidence of sequential evolution of DDT- and pyrethroid-resistant sodium channel in Aedes aegypti. PLoS Neglected Tropical Diseases, 13(6), e0007432.

Publication 2019

mMESSAGE mMACHINE high yield capped RNA kit

Austin Crna Female Molar Oocytes Ovaries Synthesis Transcription Xenopus

Corresponding Organization : Institute of Evolutionary Physiology and Biochemistry

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Variable analysis

independent variables

AaNa_v1-1 and mutants

dependent variables

Expression of AaNa_v1-1 and mutant channels

control variables

Procedures for oocyte preparation, cRNA synthesis and injection
Co-injection of AaNa_v1-1 and mutant channels cRNA with Ae. aegypti tipE cRNA in the 1:1 molar ratio

controls

Positive control: Co-injection of AaNa_v1-1 and mutant channels cRNA with Ae. aegypti tipE cRNA
Negative control: Not explicitly mentioned

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