Rabies viruses CTN strain complete genome infectious cDNA cloning, and preparation and use thereof
A rabies virus, whole genome technology, applied in the field of bioengineering, can solve the problems of primer GC% imbalance, difficult to optimize PCR reaction conditions, difficult to find annealing temperature, etc., to achieve the effect of powerful function and wide application range
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Embodiment 1
[0031] The construction method of the whole genome infectious cDNA clone of embodiment 1 rabies virus CTN strain
[0032] (1) Viruses:
[0033] The parental virus of the CTN strain rabies virus was isolated from the brain tissue of a patient infected with rabies virus in Zibo City, Shandong Province in 1953. The isolated virus was continuously passaged in the mouse brain for 56 generations, and then cultured in human diploid cells. The attenuated rabies virus vaccine strain obtained by continuing to pass 50 generations on (BMK-17) was approved by the World Health Organization (WHO) as a strain for the production of rabies vaccine in China in 1983 and 2005. This virus strain is preserved and provided by the China Institute for the Control of Pharmaceutical and Biological Products.
[0034] (1) Extraction of viral RNA:
[0035] Take a small amount of freeze-dried poisoned mouse brain, first add 0.2ml TRIzol Reagent (Invitrogen Company) to grind into a homogenate, and then add ...
Embodiment 2
[0076] Embodiment 2 The whole genome infectious cDNA clone of the rabies virus CTN strain described in the present invention is a weakly toxic animal experiment example
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