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Journal of Tea Science ›› 2007, Vol. 27 ›› Issue (2): 120-126.doi: 10.13305/j.cnki.jts.2007.02.005

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Molecular Identification, Bioinformatic Analysis and Prokaryotic Expression of the Cyclophilin Gene Full-length cDNA from Tea Plant (Camellia sinensis)

ZHANG Ya-li1,2, ZHAO Li-ping1, MA Chun-lei1,2, CHEN Liang1*   

  1. 1. Lab for Tea Germplasm, Genetic Improvement and Molecular Biology, Tea Research Institute Chinese Academy of Agricultural Sciences, Hangzhou 310008, China;
    2. Graduate School of Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Online:2007-06-25 Published:2019-09-11

Abstract: A cDNA clone, encoding cyclophilin, obtained by random sequencing of young shoot cDNA library from tea plant (Camellia sinensis). The full-length cDNA of the cyclophilin gene was 949 bp(GenBank accession No. DQ904327), containing a putative ORF of 495 bp, encoding 164 amino acids and the predicted MW were 17.47 kD and pI was 8.54, respectively. A “CAAT” signal in 5′ untranslated region and a polyadenylated signal of “AATAA”poly-A in 3′untranslated regions of cyclophilin mRNAs were found. The presumed amino acid sequences of tea plant were aligned with those of other 26 organisms through CLUSTAL W. The phylogenetic analysis based on the Neighbor-Joining method showed the similarity was greater than 85% between cyclophilin genes of tea plant and Oryza sativa (japonica cultivar-group), Solanum tuberosum, Triticum aestivum etc. Primers were designed on the open reading frame of the cyclephilin gene of tea plant to construct the expressive vector pET/Csin-Cyp. A recombinant protein about 23 kD in the Escherichia coli BL21 (DE3) was induced.

Key words: Camellia sinensis, cyclophilin, identification and analysis, prokaryotic expression

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