1.The expression, purification and identification of recombinant SARS S1 subunit expressed in E. Coli
Yan LI ; Xiaoang YANG ; Xueyuan DONG ; Yuedan WANG ; Weifeng CHEN
Chinese Journal of Immunology 1985;0(06):-
Objective:To study the course and mechanism of the immune response to SARS virus. Methods:The recombinant SARS virus S1 subunit was expressed in E. Coli according to the results of bioinformatics analysis. After purification, the recombinant S1 protein was identified by 6 serum samples of recovered SARS patients and 6 serum samples of health donors, which were collected before out-break of SARS. Results:Sequencing analysis confirmed that the recombinant protein has the same sequence of natural SARS virus S1 subunit. The recombinant S1 protein could react with all the samples from recovered SARS patients but not the control samples from healthy donors according to the results of Western blot. Conclusion:The recombinant SARS virus S1 subunit may provide a good tool for the research of immune response to SARS virus and the producing of recombinant vaccine to prevent people from SARS.
2.Mechanism study of dasatinib inhibiting phosphorylation of androgen receptor in prostatic cancer cells
Jing LIU ; Yuedan CHEN ; Jing ZHANG ; Yajie WANG ; Yong ZHANG ; Yuanbo LIU
Cancer Research and Clinic 2016;28(6):361-365
Objective To investigate the mechanism of dasatinib,tyrosine kinase inhibitor,inhibiting androgen receptor (AR) phosphorylation in prostate cancer cells.Methods HEK-293T and COS7 cell lines were cotransfected by wild-type (WT)-AR,ARY267F or ARY534F with Ack1 or Src,respectively,and Western blot was used to detect the AR phosphorylation sites.LNCaP cells were treated by EGF or heregulin without androgen,then Western blot was used to detect AR phosphorylation.After these LNCaP cells were treated by dasatinib or transfection with siRNA to silence Ack1 or Src gene,Western blot was used to observe the effect on AR phosphorylation,and quantitative real-time reverse transcription polymerase chain (RT-PCR)was applied to detect PSA mRNA and hk2 mRNA.Results After transfection,Ack1 kinase mediated the phosphorylation of AR Tyr267 in HEK-293T cells,and Src mediated AR Tyr534 phosphorylation in COS7cells.When LNCaP cells were treated by heregulin,AR Tyr267 was phosphorylated,but its phosphorylation was inhibited after these cells were treated by dasatinib or ack1 gene was silenced.When LNCaP cells were treated by EGF,AR Tyr534 was phosphorylated,but its phosphorylation was inhibited after these cells were treated by dasatinib or Src gene was silenced.EGF or heregulin raised endogenous AR target gene,PSA and hK2,mRNA levels in LNCaP cells (P < 0.05).However,after these cells were treated by dasatinib,PSA and hK2 mRNA levels induced by heregulin were decreased (P < 0.05),but those induced by EGF PSA were no significant changes (P > 0.05).Conclusion Dasatinib can inhibit AR Tyr267 and AR Tyr 534phosphorylation,and it may play a significant role in anti-prostate cancer cells by inhibiting Ack1-mediated AR Tyr-267 phosphorylation and the expression of PSA mRNA and hk2 mRNA induced by heregulin.
3. Polymodal Functionality of C. elegans OLL Neurons in Mechanosensation and Thermosensation
Yuedan FAN ; Wenjuan ZOU ; Jia LIU ; Umar AL-SHEIKH ; Hankui CHENG ; Duo DUAN ; Duo DU CHEN ; Siyan LIU ; Lijun KANG ; Yuedan FAN ; Wenjuan ZOU ; Jia LIU ; Umar AL-SHEIKH ; Hankui CHENG ; Duo DUAN ; Duo DU CHEN ; Siyan LIU ; Lijun KANG ; Luyi CHEN ; Jilei XU ; Firdosh RUHOMUTALLY
Neuroscience Bulletin 2021;37(5):611-622
Sensory modalities are important for survival but the molecular mechanisms remain challenging due to the polymodal functionality of sensory neurons. Here, we report the C. elegans outer labial lateral (OLL) sensilla sensory neurons respond to touch and cold. Mechanosensation of OLL neurons resulted in cell-autonomous mechanically-evoked Ca