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Bulletin of Botanical Research ›› 2019, Vol. 39 ›› Issue (3): 431-440.doi: 10.7525/j.issn.1673-5102.2019.03.013

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Cloning, Bioinformatics and Expression of SmSLS2 Gene in Swertia mussotii

LI Wen-Jing1, FENG Yu1, HOU Xiao-Qiang1, Sun Yan-Xiang1, Han Mei-Ling1, LI Xiao-Xue2, WANG Yong2, XIANG Bei-Bei3   

  1. 1. College of Life Science, Langfang Normal University, Langfang 065000;
    2. College of Life Science, Nankai University, Tianjin 300071;
    3. School of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, Tianjin 300193
  • Received:2018-11-13 Online:2019-05-05 Published:2019-05-11
  • Supported by:
    Colleges and Universities in Hebei Province Science and Technology Research Youth Fund(QN2018017);Tianjin Natural Science Foundation(18JCQNJC14000)

Abstract: Secologanin synthase(SLS) is the key enzyme of secoiridoid pathway. We obtained the SmSLS2 gene full-length sequence according to the transcriptome of Swertia mussotii. We cloned the gene, analyzed the bioinformation of SmSLS2, constructed the vectors and performed the gene expression. The results showed that SmSLS2 cDNA complete sequence had a length of 1 566 bp(GenBank:MH535904), encoding 521 amino acid residues, and the pI of SmSLS2 was 8.92. Domain analysis results showed that SmSLS2 had one transmembrane domain, and the protein secondary and tertiary structures were analyzed and forecasted. The gDNA sequence of SmSLS2 was 2 576 bp, contained five exons and four introns. The SmSLS2 protein shared high identity with other SLS proteins of plants. Prokaryotic expression vector pET-28a-sumo-SmSLS2 was constructed and transformed into Escherichia coli BL21(DE3) for expression under 37℃, and induced by 0.5 mmol·L-1 IPTG. The SDS-PAGE results showed that the expressed proteins were consistent with the anticipated size. SmSLS2 gene was successfully cloned from S.mussotii, and it will provide a foundation for further functional research on SmSLS2 protein and increasing the product of iridoid compound by genetic engineering in S.mussotii.

Key words: Swertia mussotii Franch, secologanin synthase2, gene cloning, bioinformations analysis, prokaryotic expression

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