Construction of plasmids containing IBDV VP2/4/3 and ChIL-18 fusion genes and co-expression genes as well as proteins expressed on Vero cells
In order to improve the immunity effect of infectious bursal disease virus (IBDV) polyprotein (VP2/4/3) DNA vaccine, fusion genes of IBDV polyprotein (VP2/4/3) gene and chicken interleukin 18 (ChIL-18) gene were constructed using the technique of splicing by overlapping extension (SOEing) and three...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2010-01-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.01.001 |
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| Summary: | In order to improve the immunity effect of infectious bursal disease virus (IBDV) polyprotein (VP2/4/3) DNA vaccine, fusion genes of IBDV polyprotein (VP2/4/3) gene and chicken interleukin 18 (ChIL-18) gene were constructed using the technique of splicing by overlapping extension (SOEing) and three times PCR. These fusion gene fragments were directly cloned into pCI, an eukaryotic expression vector, to produce recombinant expression plasmids pCI-VP2/4/3-ChIL-18 and pCI-ChIL-18-VP2/4/3. Then, co-expressing plasmid co-pCI-VP2/4/3-ChIL-18 was constructed on the basis of pCI-VP2/4/3 and pCI-ChIL-18 constructed in our lab by PCR. These recombination plasmids were transfected into Vero cells introduced by Lipofectamine 2000 reagent. The interest proteins were detected by indirect immunofluorescence, indicating that these fusion genes were correctly expressed in Vero cells. It is provided a basis for the research of immune-enhancement of ChIL-18 and manufacturing highly effective and inexpensive DNA vaccination against IBDV. |
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| ISSN: | 1008-9209 2097-5155 |