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Establishment ofChlamydomonas reinhardtii chloroplast expression

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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Author information

Correspondence to Guifang Shen.

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Cite this article

Fan, G., Su, N., Zhang, Z. et al. Establishment ofChlamydomonas reinhardtii chloroplast expression. Chin.Sci.Bull. 44, 1659–1664 (1999). https://doi.org/10.1007/BF03183484

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Keywords

  • HAV-VP3P1 gene
  • HCV-C gene
  • C. reinhardtii
  • chloroplast expression system
  • homologous recombinant