Method for constructing interference plasmid of human fibrinogen hf12 shRNA, and pharmic usage
A technology of cellulose intermediary and construction method, applied in the field of bioengineering, can solve the problem of no definite treatment method for severe hepatitis
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[0042] Below in conjunction with specific embodiment the present invention is described in further detail:
[0043] The present invention proposes the method for constructing hfgl2 shRNA interference plasmid and confirms its interference effect at cellular level and comprises the following steps:
[0044] 1. The construction part of the carrier
[0045]Method for constructing hfgl2 shRNA interference plasmid:
[0046] 1. Screen the target sequence according to the hfgl2 prothrombinase cDNA sequence and design principles, design the hfgl2 shRNA template according to the target sequence, then design the template in the downstream primers, amplify the U6 promoter by PCR, and introduce restriction sites at both ends BamH I and Hind III (see Figure 1 for the two target sequences of hfgl2 and the upstream primers of the U6 promoter).
[0047] 2. Insert the U6 promoter with the hfgl2 shRNA template downstream into the pMSCVneo vector (Figure 2) corresponding restriction sites BamH ...
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