Construction of 21ShRNA fluorescent expression vector of long-term silencing esophageal cancer cell HIF-1Alpha gene
A technology of HIF-1 and expression vector, applied in the field of biomedicine, can solve problems such as decreased PDT effect
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[0022] The construction of a 21ShRNA fluorescent expression vector for long-term silencing of esophageal cancer cell HIF-1α gene according to the present invention comprises the following steps:
[0023] (1) Design of HIF-1α interference sequence
[0024] The HIF-1α gene ID and mRNA full-length sequence number were obtained from NCBI (NM_001530.3), and the WHITEHOUSE software was used to predict and analyze the interference sequence consisting of 21 nucleotides, and the GC content was controlled at 30%-50%; at the same time, the s iDirect online design software, input the full length of the obtained mRNA sequence into the sequence box to be predicted, select the design policy as the Reynolds equation, control the GC content at 30%-50%, run the remote prediction program, and obtain the prediction result;
[0025] (2) BLAST analysis
[0026] Using NCBI's Nucleotide Blast to compare the predicted nucleotide sequences, design a 21-nucleotide HIF-1α interference sequence, the sens...
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