Establishment ofChlamydomonas reinhardtii chloroplast expression |
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Authors: | Guochang Fan Ning Su Zhonglin Zhang Xiangfu Wu Guifang Shen |
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Institution: | FAN Guochang, SU Ming, ZHANG Zhonglin, WU Xiangfu and SHEN Guifang
1. Bio-Tech Research Centre , Chinese Academy of Agricultural Sciences , Beijing 100081 , China ;
2. Beijing Institute of Radiation Medicine, Beijing 100850, China ;
3. Shanghai Institute of Biochemistry , Chinese Academy of Sciences , Shanghai 200031 , China Corresponding author |
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Abstract: | An expression vector pAC III containing the chimeric gene HAV-VP3P1 and HCV-C gene has been constructed and transferred toC. reinhardtii by the biolistic method. The transformants have been identified by PCR, Southern-blotting, Northern-blotting and Western-blotting assays after selecting on resistant medium and incubating in the dark. The results show that the chimeric gene has replaced thechlL gene ofC. reinhardtii chloroplast genome and expressed correctly under the control of theC. reinhardtii chloroplast double promoters 5′chlL-5′atpA. The analysis of SDS-PAGE indicates that the expressed protein accounts for 5.31 % of total soluble protein. |
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Keywords: | HAV-VP3P1 gene HCV-C gene C reinhardtii chloroplast expression system homologous recombinant |
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