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METHOD OF INTRODUCING DOUBLE-STRANDED DNA INTO THE BODY OF KERRIA CHINENSIS
METHOD OF INTRODUCING DOUBLE-STRANDED DNA INTO THE BODY OF KERRIA CHINENSIS
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机译：将双链DNA导入中国沙棘的方法
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#$%^&*AU2020100222A420200326.pdf#####22 ABSTRACT A method of introducing double-stranded RNA into the body of kerria chinensis includes the steps of: extracting the total RNA of kerria chinensis, reversely transcribing the RNA into cDNA, performing a PCR amplification, amplifying the product and a L4440 carrier by Sacd and SalI double enzyme digestion, the enzyme digestion product using T4 ligase for connection overnight, transferring the obtained FAD-L4440 recombinant plasmid into HT115 competent cells, amplifying and culturing the broth until OD 6 0 =0.1~0.7, adding the broth into IPTG for an induction for 2~5h to obtain FAD-L4440-HT115 thallus, diluting the thallus, and smearing the thallus directly onto the kerria chinensis larva. This method effectively interferes the FAD gene expression to transfect dsRNA into the body of kerria for effective interference, so as to reduce the individual secretion volume and provide references of insects whenever transfection methods such as injection methods and feeding which are inapplicable in the RNAi transfection process.FAD gene 5GAGCTC GTCGAC 3' 3'CTCGAG CAGCTG5 Sac5 L4440 Sail SacI + sacl Double Enzyme Digestion T4 Ligase Plasmid Extraction FAD+L4440 IPTG Induction 1500 C FIG. 1 500bp 2sobp 750bp lS0bp 250bp 2S0bp A B C FIG. 2
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