Recombinant adenovirus for preventing baby's diarrhea
A technology for recombinant adenovirus and infant diarrhea, applied in the field of recombinant adenovirus, can solve problems such as economic loss, and achieve the effects of ensuring safety in use, solving the problem of virus mutation, and being easy to prepare
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[0025] Cloning of Rotavirus VP6 Gene
[0026] 1. Diarrhea and stool samples were collected from the pediatric outpatient clinic of a domestic hospital, which was confirmed to be rotavirus infection by electron microscopy. Rotavirus genomic RNA was extracted from this sample, and purified as a template. According to the known VP6 gene nucleotide sequence, the upstream and downstream primers were designed, with EcoRI and BamHI restriction sites, respectively, and reverse transcription PCR was used. (RT-PCR) method to amplify the VP6 full-length gene to obtain a fragment with a length of about 1.4Kb. The PCR amplified product was recovered by the low-melting point agarose method, digested by EcoR1 and BamHI, and inserted into the corresponding restriction site of pGEM-7zf vector, and the positive clone pGEM-VP6 inserted into the rotavirus gene was screened out. The sequence analysis results of the positive clones proved that the cloned VP6 gene was completely correct.
[0027] ...
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