Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (2): 1-9.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0498
YANG Jia-hui(), SUN Yu-ping, LU Ya-ning, LIU huan, LU Cun-fu(), CHEN Yu-zhen()
Received:
2021-04-15
Online:
2022-02-26
Published:
2022-03-09
Contact:
LU Cun-fu,CHEN Yu-zhen
E-mail:yangjiahui940529@163.com;lucunfu@bjfu.edu.cn;chenyuzhen@bjfu.edu.cn
YANG Jia-hui, SUN Yu-ping, LU Ya-ning, LIU huan, LU Cun-fu, CHEN Yu-zhen. Abiotic Stress Resistance of Escherichia coli Transformed with Arabidopsis thaliana AtTERT Gene[J]. Biotechnology Bulletin, 2022, 38(2): 1-9.
Fig. 1 PCR amplification of AtTERT gene A:AtTERT gene with pET32a vector sequence. B:AtTERT gene with pGEX-4T-1 vector sequence. M:5 000 bp DNA marker
Fig.2 Double digestion identification of pET32a/ pGEX-4T-1-AtTERT A:Identification of pET32a-AtTERT recombinant plasmid after enzyme digestion. B:Identification of pGEX-4T-1-AtTERT recombinant plasmid after enzyme digestion. M:5 000 bp marker. 1,2:Recombinant plasmid after enzyme digestion. 3:Control plasmid
Fig. 3 Detection of GST-TERT protein by SDS-PAGE A:Expression of soluble protein under different IPTG concentrations at 20℃. B:Expression of soluble protein under different IPTG concentrations at 30℃. C:Expression of inclusion body protein under different IPTG concentrations at 20℃. D:Expression of inclusion body protein under different IPTG concentrations at 30℃. M:180 kD marker
Fig. 4 Western blotting identification of GST-AtTERT fusion protein A:Identification of GST-AtTERT fusion protein pattern(1,2:bacterial protein induced by empty vector control bacteria and transgenic recombinant bacteria;3,4:soluble protein induced by empty vector control bacteria and transgenic recombinant bacteria;5,6:inclusion body protein induced by empty vector control bacteria and transgenic recombinant bacteria). B:Identification of purified GST-AtTERT fusion protein(1,2:soluble protein induced by empty vector control bacteria and transgenic recombinant bacteria;3:outflow after incubation;4:outflow after miscellaneous protein washing out;5-7:protein after elution)
Fig.5 Effect of AtTERT on the growth characteristics of E.coli **indicates that the difference is extremely significant at P<0.01;*indicates that the difference is significant at P<0.05. The same below
Fig.6 Effect of AtTERT on the survival rate of E. coli under salt stress A:Growth status of empty control bacteria(pET32a)and transgenic recombinant bacteria(pET32a-AtTERT). B:Survival rate of E.coli when the bacterial solution diluted 104 times
Fig.7 Effect of AtTERT on the survival rate of E. coli under mannitol stress A:Growth status of empty control bacteria(pET32a)and transgenic recombinant bacteria(pET32a-AtTERT)on the LB solid medium containing mannitol. B:Survival rate of E.coli when the bacterial solution diluted 104 times
Fig.8 Effect of AtTERT on the survival rate of E. coli under cold stress A:Growth status of empty control bacteria(pET32a)and transgenic recombinant bacteria(pET32a-AtTERT)on the LB solid medium after repeated freezing and thawing in liquid nitrogen. B:Survival rate of E.coli when the bacterial solution diluted 104 times
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