[Expression of c-terminal of Parkin in E. coli and the preparation of antiserum].

To clone and express 3'-terminal of Parkin gene in E. coli, and prepare its antiserum for further study.The glutathion-sulfate-transferase (GST) fusion expression plasmid of 3'-terminal of Parkin gene (937-1959 bp) was constructed and transferred to JM 105. After being treated with Triton-100 (1%) and Tween-20 (1%) and purified with affinity chromatograph, GST-Parkin C was used to immunize New Zealand rabbits to acquire antiserum. Antiserum was analysed with immunoblot.The GST-Parkin C protein was expressed in JM 105, existing in the form of inclusion body with a molecular weight of around 42 kD; The purity of GST-Parkin C was up to 95%; the titer of antiserum was 1:64; Immunoblotting showed that the prepared antiserum could react specifically with 51.6 kD protein extracted from the mouse brain.A high level of expression of GST-Parkin C is obtained in JM 105, and its antiserum can be prepared successfully.