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Fig. 5. RNAi-mediated knockdown of Aaslif and AaiCAT2 inhibits
amino acid stimulation of the vg gene. 1 day-old mosquitoes were
injected with 0.6-1.0 µg of the following dsRNAs: the non-coding region of
a control bacterial gene (MAL), the coding region of neutral amino acid
transporter (NAT), the coding region of the Aaslif gene (Slif), the
coding region of the AaiCAT2 gene (iCAT2), or a 1:1 mixture of both
Aaslif and AaiCAT2 dsRNAs. Mosquitoes were allowed to
recover for 5 days. Fat bodies from these mosquitoes were then dissected and
cultured in either the presence or absence of amino acids (AAs) for 6 h. Total
RNA was isolated from three groups of six fat bodies per treatment. cDNA was
synthesized from equal amounts of DNase I-treated total RNA. (A) Gene
expression was analyzed using vg-specific real-time PCR primers. Data
were normalized by real-time PCR analysis of actin levels in the cDNA
samples. Values are means ± s.e.m. of triplicate samples. (B) Knockdown
of transporter genes was confirmed by RT-PCR analysis of the same cDNA used
for the analysis in A.
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