Molecular Cloning of Cold-induced JcDnaJ20p Promoter from Jatropha curcas and Its Functional Identification in Transgenic Tobacco

doi:10.13560/j.cnki.biotech.bull.1985.2020-0620

Abstract

Abstract:

DnaJ20 is a cold-induced gene based on Jatropha curcas cold transcriptome data. In order to identify the cold-induced activity of this gene,the promoter sequence of DnaJ20 gene（JcDna20）from the genomic DNA of Jatropha curcas leaf was cloned based on PCR technology,and designated as JcDnaJ20p.,then,a plant binary expression vector pCambia1381Z-JcDnaJ20p-GUS was constructed by fusing JcDnaJ20p promoter with GUS reporter gene,and transferred into tobacco（Nicotiana tabacum）by Agrobacterium tumefaciens system. The results showed that the sequence length of DnaJ20 gene promoter was 2 023 bp. Sequence analysis found that JcDnaJ20p had the typical eukaryotic promoter core regions（TATA-box and CAAT box）. In addition,hormonal responsive elements（abscisic acid and gibberellin）and resistance-related responsive elements（low temperature and drought）were identified in the sequence of JcDnaj20p. Compared to plants transformed with empty vector of pCambia1381Z-GUS and pCambia1381Z-35S-GUS,histochemical staining of the transgenic tobacco with pCambia1381Z-JcDnaJ20p-GUS showed that the expression of GUS gene increased under 15 and 4℃ cold treatments for 24 h,indicating that JcDnaJ20p was an efficient cold-inducible promoter.

Key words: Jatropha curcas, cold induction, promoter, DnaJ20, gene cloning, functional identification

WANG Hai-bo, GUO Jun-yun. Molecular Cloning of Cold-induced JcDnaJ20p Promoter from Jatropha curcas and Its Functional Identification in Transgenic Tobacco[J]. Biotechnology Bulletin, 2021, 37(2): 24-31.

Figures/Tables 4

References 30

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