A nucleic acid sequence that can be used for molecular pest control and its application
A nucleic acid and sequence technology is applied in the application field of preventing and controlling Spodoptera litura, and can solve problems such as water pollution, pest generation, drug resistance and the like
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Embodiment 1
[0051] 1.4E specific primer design
[0052] Transcriptome data from Spodoptera litura After Parasitization by Microplitis bicoloratus.PLoS ONE 9( 10):e110967.doi:10.1371 / journal.pone.0110967), the 4E gene sequence was selected, the size of the 4E gene sequence (as shown in SEQ ID No.1) was 633bp, and a total of 210 amino acids were encoded (SEQ ID No.2). Specific primers were designed using the primer design software Primer Premier 5. Upstream primer L4440-4E-F (SEQ ID No. 3) and downstream primer L4440-4E-R (SEQ ID No. 4).
[0053] 2. Extraction of total RNA and cDNA synthesis of Spodoptera litura larvae
[0054] In this experiment, the laboratory protocol was used to extract total RNA from Spodoptera litura larvae. The specific steps are as follows:
[0055] 1) Take 10 3-day-old larvae, place them in EP tubes with a small amount of RNAisoTM Plus lysate, mash and grind them thoroughly, continue to add RNAisoTM Plus lysate until the total volume reaches 1ml, vortex and mix ...
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