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Expression of annfaf Gene Expressed Specifically in Strawberry
Fruit in E.coli and Preparation of Its Antiserum |
| Wang Guanlin1 Xia Ran1 Fang Hong-Jun1 Yang Huaiyi2 Na Jie1 |
| (1.College of Life Science, Liaoning Normal University, Dalian 116029, China; 2. Institute of Microbiology, Chinese Academy of Science, Beijing 100080, China) |
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Abstract Abstract: The fusion expression vector and the non-fusion expression vector were constructed by the open reading frame (ORF) of annfaf expressed specifically in strawberry (Fragaria ananassa Duch.)fruit subcloning to plasmid pET-30a. The recombinant vectors were transformed and expressed high-effectually in E.coli BL21(DE3), and the target protein was detected by SDS-PAGE with a molecular weight of 35 kD. The SDS-PAGE also indicated that expression product was inclusion body. The study of effective expression showed that the optimum culture conditions of E.coli were 37 ℃ for 4 h, and the target protein content of the total expression proteins was increased clearly after adding rifamycin in the medium. The target protein recovered from SDS-PAGE was used to immunize rabbit, and antiserum with high titer was obtained. The ELISA and Western bloting analysis indicated that the reaction between antiserum and antigen was highly specific and effectual. The experement results wear a foundation of the study on function and location of annafaf protein in the cells of strawberry fruit.
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Received: 01 January 1900
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