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| Prokaryotic Expression and Activity Detection of Ad5/Ad35 Fiber Gene |
| ZHANG Wen-feng1,2, ZHANG Qiong-yu3, BO Hua-ben1,2, SHAO Hong-wei1,2, LI Xiao-cheng1,2, WANG Teng1,2, HUANG Shu-lin1,2 |
1. School of Life Science & Biopharmacology, Guang Dong Pharmaceutical University, Guanzhou 510006, China;
2. Guang Dong Provincial Key Laboratory of Biotechnology Candidate Drug Research, Guangzhou 510006, China;
3. Department of Basic Medical Science, Yongzhou Vocational Technical College, Yongzhou 425100, China |
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Abstract Aim: To construct the prokaryotic expression vector for Ad5 fiber and Ad35 fiber genes, purify proteins and study the activity. Methods: Recombinant plasmid pET28a-Ad5 fiber and pET28a-Ad35 fiber were transformed into E.coli BL21(DE3) and the proteins expression were induced with IPTG. The expressed proteins Ad5 fiber and Ad35 fiber were purified by Ni2+ affinity chromatography and identified by SDS-PAGE. The bioactivity of proteins Ad5 fiber and Ad35 fiber were certificated by competitive binding method. Results: The proteins Ad5 fiber and Ad35 fiber can be detected in the supernatant. The proteins Ad5 fiber specifically inhibited the infectivity of the Ad5 virus in a dose-dependent manner, while had no effect on the Ad5F35-mediated gene transfer. The proteins Ad35 fiber blocked the Ad5F35-mediated gene transfer in a dose-dependent manner and had no effect on the infectivity of the Ad5 virus in competition experiments. Conclusion: Recombinant vectors Ad5 fiber and Ad35 fiber were constructed successfully. The proteins Ad5 fiber and Ad35 fiber could specifically bind with different receptors. The study will lay a foundation for the investigation on the mechanism of Ad5F35 infection.
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Received: 06 September 2013
Published: 25 December 2013
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