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1. What is the baculovirus?
Baculoviruses are arthropod-specific viruses containing large double-stranded circular DNA genomes of 80,000–180,000?bp. In recent years, they have been utilized for producing complex eukaryotic proteins in insect cell cultures.
2. What do the P1 and P2 mean?
P1 is the code of recombinant virus which has packaged foreign target gene. P1 stock usually has a low titer (106 ~ 107pfu). While, P2 is the code of virus amplified from P1. P2 stock usually has a high titer (>107pfu).
3. What are the advantages of using baculovirus to express target proteins?
There are several benefits of using baculovirus to express target proteins, such as larger inserts, ability of post-translational modifications, enhanced protein folding and function, high expression levels and safety.
4. How long does it take to generate a new recombinant baculovirus?
Creative Biogene can provide the fastest turnaround time of any supplier in the industry. But the exact lead time depends on your detailed information and request of the project.
6. Is there a packaging limit for the baculovirus?
The baculovirus rod will continue to elongate as required to package the DNA. Thus, the system could theoretically accommodate hundreds of Kb. But, the standard cloning techniques will limit the insert size. It may become an issue before packaging limits.
7. What could be the cause for degrading of bacmid DNA and what recommendations do you have to fix this?
There are two possible reasons cause for it. Firstly, DNA stored improperly. Ensure that purified bacmid DNA is stored at -20 degrees in aliquots to avoid repeated free/thaws. Secondly, High molecular weight bacmid DNA handled improperly: When isolating bacmid DNA, do not vortex the DNA solution; additionally, do not resuspend DNA pellets mechanically.
8. Can a P2 viral stock be used to generate more viruses? How about a P3 or P4 stock?
Yes, the same protocol used to make your P2 viral stock can be used to make a P3, P4, or P5 viral stock. We don’t recommend making the stock higher than P5, as more defective interfering particles will be produced.