Recombinant HBV (Hepatitis B Virus) vector for expressing specific HBVshRNA (Hepatitis B Virus Short Hairpin Ribonucleic Acid) as well as construction method and application of vector
A specific, carrier technology, applied in the field of molecular biology, which can solve the problems of inability to synthesize replication and complete inhibition.
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[0027] In order to make the object, technical solution and advantages of the present invention clearer, preferred embodiments of the present invention are described in detail below. The experimental methods for which specific conditions are not indicated in the preferred embodiments are usually carried out according to conventional conditions, or according to the conditions suggested by the manufacturer.
[0028] The LJ196 and LJ96 plasmids used in the present invention were donated by Professor Daniel D. Loeb, and are described in the published literature (Liu N, Ji L, Maguire ML, Loeb DD. cis-Acting sequences that contribute to the synthesis of relaxed- circular DNA of human hepatitis B virus. J Virol. Jan 2004; 78(2): 642-649.).
[0029] 1. Deletion mutant rHBV construction process
[0030] On the basis of the plasmid LJ196 (SEQ ID NO.1) that does not express any viral protein, the DNA fragment of the LJ196 plasmid is deleted by reverse PCR technology, so that the RC DNA...
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