| Plant activator protein from Alternaria sp., which is short for ActP, is a kind of thermostable protein, which can promote plant growth. In this study, we identified the activator protein from Alternaria sp., made antibodies against ActP, screened of the expression library of Alternaria sp. by immunological methods, obtained the gene of ActP and purified the expression protein.The activator protein from Alternaria sp. was separated by 2D-PAGE. Useing 2mM EDTA solution separated the ActP protein with different method of isoelectricfocusing (IEF) and two-dimensional gel electrophoresis. The protein was separated by 2D-PAGE, and confirms its pI as 4.Specificity of the antibodies against the 35kD ActP35 protein was purified successfully. The antigen protein was the 35kD activator protein ActP, which is short for ActP35. The antiserum was obtained from the rabbit blood. But the antisera strongly cross react to E.coli, so it is unable to use for screening the expression library directly. In this study, the method of blot-affinity purification was improved and specific antibodies which didn't react to E.coli were obtained. It is a new way to obtain specific antibodies simply.The expression library of Alternaria sp. was screened perfectly useing the antibodies against Actp35. 9 positive clones were obtained and sequenced. Comparing with the amino acid coding of activator ActP35, and make sure that one of them is right clone, which is short for actd-2. The actd-2 gene was expressed in E.coli and the fusion protein was purifird by metal-ion affinity chromatography.The interact protein in the Arabidopsis thalian of ActP35 was detected by far Western , immunoaffinity chromatography. We find a kind of antibody which could specifically react with ActP35 and the Rubisco of Arabidopsis thaliana, the two proteins may have the same sequence. Maybe it is the evidence for the fact that the follow-up study plant activator protein and promotes the plant photosynthesis.
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