Cloning and Prokaryotic Expression of Phosphoglucomutase Genes cPGM and pPGM from Jatropha curcas

doi:10.7668/hbnxb.20192060

Abstract

Abstract: In order to explore the function of phosphoglucomutase(PGM) in the metabolism of sucrose and starch in plants. Based on homologous sequence alignment, a cytoplasmic PGM gene(named JccPGM) and a chloroplastic PGM gene(named JcpPGM) were identified from the Jatropha curcas genome. The expression levels of JccPGM and JcpPGM in different organs and under chilling stress were detected by qRT-PCR method. The prokaryotic expression recombinant vectors of pGEX-4T-1-JccPGM and pGEX-4T-1-JcpPGM were constructed, and then fusion proteins were induced in Escherichia coli BL21(DE3) strains. The results showed that JccPGM and JcpPGM encoded proteins of 582, 637 aa, respectively. Cluster analysis showed that pPGM contained a chloroplast localization signal peptide at the N-terminal, whereas cPGM had four more peptide sequences of -¹⁰⁸VGVDGS¹¹³-, -¹⁸³SGPE¹⁸⁶-, -²⁸³GKSNSE²⁸⁸-, -⁴⁷⁰SLGEVN⁴⁷⁵- than pPGM. qRT-PCR analysis revealed that cPGM and pPGM expressed specifically in different organs, abundantly in leaves, but scarcely in roots and seeds. Escherichia coli BL21(DE3) induction and SDS-PAGE analysis indicated that the fusion protein molecular weights were 90.7, 97.0 ku, respectively, which was consistent with the predicted weights. In conclusion, this study lays a foundation for further studies on the protein functions of cPGM and pPGM and the mechanisms underlying sucrose and starch accumulation and stress responses in J.curcas.

Key words: Jatropha curcas L., Phosphoglucomutase, Gene cloning, Prokaryotic expression

CLC Number:

Q78
S759.03

WANG Haibo, LI Furong, YANG Jincui, GAO Yong, GUO Junyun. Cloning and Prokaryotic Expression of Phosphoglucomutase Genes cPGM and pPGM from Jatropha curcas[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(4): 23-30. doi: 10.7668/hbnxb.20192060.

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