Adenovirus Packaging Experiment Service
First Edition (Revised on April, 2016)
By means of virosome as a gene vector, use the infectivity of virus, spontaneous infection of mammalian cells. Introducing target genes into mammalian cells, thus achieve excessive expression or interference of the target product. The adenovirus expression system employed an artificially modified Ad5 virus system, usually included one expression plasmid and one to more helper plasmids. It used 293 cells as hosts to breed viruses,by centrifugation ultrafiltration and other processes to prepare and enrich recombinant adenovirus particles, to complete the packaging process of adenovirus. Packaged recombinant virus infected cells, the target gene was free from the genome of the host cell and independent expression, could be realized the target protein expressed rapidly and high abundance. At the same time, avoid genetic mutations caused by genetic integration.
[EXPERIMENTAL PROCEDURES AND PERIOD]
Determine the experiment scheme
1. Analyze the customer information
2. Determine the experiment scheme
Acquire target gene
1. Obtain the PCR target gene fragment
2. Chemical synthetic gene
1. Construct vector
2. Extract plasmids and treated to remove endotoxin
1. Transfect the expressed plasmid and the helper plasmid into 293 cells
2. Cell culture
Collect recombinant virus
1. Collect, concentrate, repack and preservate the recombinant viruses
2. Detect the recombinant viral titers
[CUSTOMER SUPPLIED INFORMATION]
1. Detailed information on the DNA sequences of target genes and their related fragments.
2. Clients' specific requirements.
[RESULTS PROVIDED TO THE CUSTOMER]
1. Experiment schemes.
2. Gene clone positive plasmids.
3. The viral titers ranged greater than or equal to 1*1011~1*1013TU/ml in the recombinant virus reservoir.