RNA (Ribonucleic Acid) interference fragment of donkey Zfy gene, expression vector and application of RNA interference fragment
A technology of RNA interference and expression vectors, applied in the field of molecular genetics, can solve problems such as difficult control, large randomness, and instability, and achieve low-cost effects
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[0015] The technical solutions provided by the present invention will be described in detail below in conjunction with the examples, but they should not be interpreted as limiting the protection scope of the present invention.
[0016] 1. Construction of the RNA interference fragment and expression vector of the donkey Zfy gene
[0017] This application has screened 5 pairs of siRNA fragments through preliminary work, and then according to the requirements of the vector and the promoter, add restriction sites, loops and termination sequences to the 5' and 3' of each siRNA fragment, and the final form The shRNA oligonucleotide sequence is: T+sense strand 19nt target sequence+stem-loop structure (TTCAAGAGA)+target sequence reverse complement sequence+RNAPolyIII polymerase transcription termination site (TTTTTT)+XhoI restriction site (GAGCT) ( Entrust Shanghai Biological Engineering Co., Ltd. to synthesize the shRNA oligonucleotide sequence), as shown in Table 1.
[0018] Table ...
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