Cloning and Prokaryotic Expression of ORFV <i>115</i> Gene

LIN Yusheng; HUANG Lili; JIANG Jinxiu; ZHANG Jingpeng; LIU Weiwei; LIU Qinghua; HU Qilin

doi:10.19303/j.issn.1008-0384.2022.007.005

Volume 37 Issue 7

Jul. 2022

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Article Navigation > Fujian Journal of Agricultural Sciences > 2022 > 37(7): 850-854

LIN Y S, HUANG L L, JIANG J X, et al. Cloning and Prokaryotic Expression of ORFV 115 Gene [J]. Fujian Journal of Agricultural Sciences，2022，37（7）：850−854 doi: 10.19303/j.issn.1008-0384.2022.007.005

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LIN Y S, HUANG L L, JIANG J X, et al. Cloning and Prokaryotic Expression of ORFV 115 Gene [J]. Fujian Journal of Agricultural Sciences，2022，37（7）：850−854 doi: 10.19303/j.issn.1008-0384.2022.007.005

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Cloning and Prokaryotic Expression of ORFV 115 Gene

doi: 10.19303/j.issn.1008-0384.2022.007.005

1.
Institute of Animal Husbandry & Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350013, China
2.
No. 5 Middle School, 11^th Division, Xinjiang Production and Construction Corps, Urumqi, Xinjiang　830000, China
3.
College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China

Received Date: 2022-02-18
Rev Recd Date: 2022-04-20

Available Online: 2022-08-07

Publish Date: 2022-07-28

Abstract

Abstract

Objective ORFV 115 was cloned for a study on the gene. Method Oligo 7 software was used to design and select primers for specific amplification of the gene. Using the genome of ORFV FJ-ND as template, the sequence was obtained by PCR. The gene was then cloned into pGEX-6p-1 to obtain the recombinant plasmid pGEX-6P-115 for sequencing and identification prior to transformation to RosettaganmiB (DE3) receptor cells. Expression conditions of the recombinant fusion protein were optimized with respect to IPTG concentration and induction time. SDS-PAGE was used to analyze the expressed target protein. Result ORFV 115 was 450 bp encoded 149 amino acids and could be expressed in RosettaganmiB. It was approximately 42 kDa in molecular weight and mainly expressed as inclusion body. Conclusion ORFV 115 was successfully cloned, the pGEX-6P-115 prokaryotic expression plasmid constructed, the recombinant protein purified, and the expression conditions optimized to facilitate further studies on the mechanism and role it plays in the infection on host animals.
- Orf virus,
- ORFV 115,
- cloning,
- prokaryotic expression

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References(16)

References

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