Identification and Prokaryotic Expression of the TRX-y1-1 Gene from Cardamine hupingshanensis
GAO Lilong
LI Yao
LUO Guoqiang
ZHOU Yifeng
Abstract：To elucidate the antioxidant mechanisms of the hyper-selenium-accumulating plant Cardamine hupingshanensis under selenium stress,bioinformatics methods were used to identify and analyze the thioredoxin-y(TRX-y)gene family.Five members of the TRX-y gene family were identified in C.hupingshanensis,distributed across four chromosomes.All members contained the conserved active center"WCGPC"and were localized in chloroplasts.Phylogenetic analysis revealed that they clustered in Class I with AtTRX-y from Arabidopsis thaliana,and the ChTRX-y isoform possesses a specific motif.Selenium stress expression profiling revealed that high selenium concentration(80000μg/L)significantly upregulated the expression of ChTRX-y1-1 and ChTRX-y1-2 in roots(24 hours)and leaves(9 hours),suggesting that these were core selenium-responsive genes.ChTRX-y1-1 was further cloned,and the pET28b-ChTRX-y1-1 recombinant vector was constructed.Expression in Escherichia coli BL21 was achieved using induction conditions of a temperature of 25℃,a molar concentration of 0.1 mmol/L Isopropyl-beta-D-thiogalactopyranoside and a duration of 10 h.A highly pure recombinant protein of approximately 59 kDa was successfully obtained,reaching a concentration of 1.26 mg/mL after ultrafiltration.This study revealed the key role of ChTRX-y1-1 in selenium stress and established a prokaryotic expression system for its expression,providing a molecular basis for further investigation into the selenium metabolism mechanism in C.hupingshanensis.
Keywords：Cardamine hupingshanensisthioredoxinselenium stressgene identificationprokaryotic expression
Publication Date：2025-12-30
Online Publishing Date：2025-12-10(First online date of this platform, not the publication date of the document)
Pages：7( 476-482 )