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《Chinese Journal of Microecology》 2017-08
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The cloning,expression and antitumor activity of Unigene1245 transcript from Lentinula edodes C_(91-3)

SHAO Liqun;LUAN Hangqi;WANG Yu;ZHANG Wei;ZHONG Mintao;WANG Xiaoli;LI Xingyun;SHA Li;HUANG Min;Department of Medical Microbiology,Dalian Medical University;  
Objective To clone the Unigene1245 of Lentinula edodes C_(91-3)and explore the antitumor activity of the target protein.Methods The total RNA was extracted from the mycelium of Lentinula deodes C_(91-3)to synthesize the cDNA by using reverse transcription.Unigene1245 gene coding sequence was obtained via RACE technology,then bioinformatics analysis was performed by using NCBI.The gene was amplified by using PCR with the specific primers,and connected with pET-32a(+)vector.The recombined vector was transformed into E.coli Rosetta-gami(DE3),and harvested by inducing,purifying and refolding.CCK-8method was used to prove its anti-tumor activity.Results Lentinula edodes C_(91-3)transcript Unigene1245 was found to be a member of YjgF/YER057c/UK114 family.The recombinant expression plasmid was constructed successfully and the target protein was successfully induced.The results of CCK-8showed that the protein had a certain inhibitory effect on the proliferation of HepG2 cells,which had certain time and concentration dependence.Conclusion Unigene1245 was successfully cloned and induced,which was found to have the ability to inhibit the proliferation of HepG2 cells.This result laid the foundation for subsequent study to explore the antitumor mechanism.
【Fund】: 国家自然科学基金(501579)
【CateGory Index】: R285;S646.12
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