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Virologica Sinica, 19 (2) : 141-145, 2004
Research Article
Study on Heredity Stability of a Recombinant Baculovirus Expressing Human Interleukin-12 in Sf9 Cells during Serial and Undiluted Passages
Institute of Virology, Wuhan University, Wuhan 430072, China
 Correspondence:
(256.33KB)  
Abstract
The strain purified from a baculovirus expressing recombinant human interleukin-12 by plaque selection was serially and undilutedly passaged up to 55th generation in Spodoptera frugiperda 9(Sf9) cells. Intracellular viral (ICV) DNAs were extracted from the infected Sf9 cells of P15, P25, P35, P45, P55 generation of recombinant baculovirus. A 2.0kb DNA fragment including sequences of P35 cDNA, polyhedrin promoter, P10 promoter and P40 cDNA was amplified by PCR. The sequence analysis indicates that there is no mutation in 2.0kb nucleotide sequence during the P15 to P35 passages. However, the point mutation was detected at three nucleotide residue sites in P35 cDNA sequence (461T→C,517A→G and 630C→T) and one nucleotide “T” was inserted between +1 position of polyhedrin promoter and the upstream of BamH I recoganizing site at P45 generation. When passaging to P55, the insertional mutation (-136T-135), deleton mutation (-122T) and point mutation (-168G→T) happened in the P10 promoter cassette besides the above mutations. These results show that serial and undiluted passaging of engineering baculovirus can result in foreign gene mutation.
  Published online: 20 Apr 2004
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