Mink enteritis parvovirus whole genome infectious clone and construction method and application thereof
An enteritis parvovirus, infectious cloning technology, applied in applications, viruses, genetic engineering, etc., can solve the problems of few people's research, difficulty in cloning full-length genome, affecting the establishment of reverse genetics operation platform, etc., to overcome the capacity Small, simple method, high accuracy effect
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[0030] In order to better understand the present invention, the present invention will be further described below in conjunction with specific biological materials and parameters, but the present invention is not further limited. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art; all the reagents involved can be obtained from commercial sources.
[0031] 1. Transformation of vector pUC18
[0032]First linearize pUC18 with Hind Ⅲ, perform 1% agarose gel electrophoresis at 37°C for 1 hour, cut out the DNA of the target band, recover and purify it; then use Blunting blunting enzyme to treat the purified vector, and act at 37°C for 10 minutes 1% agarose gel electrophoresis at 70°C for 5 minutes, and the DNA of the target band was excised, recovered and purified. This step changed the two ends of the vector fragment from sticky ends to blunt ends, and named it pUC18-M; The vector purified in the previ...
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