Preliminary research on the expression, purification and function of the apoptotic fusion protein, Sival.
The objective of the present study was to investigate cloning, expression, and functions of the recombinant protein, Siva1. Siva1 gene was synthesized by RT-PCR from HCT116 cells. Plasmids were cleaved with the restriction endonuclease, BamH1/Sal1 and products were connected to pQE30, which underwent cleavage by BamH1/Sal1. The recombinant plasmid, pQE30-Siva1, was identified after digestion with restriction endonucleases followed by transformation into E. coli M15. Expression of Siva1 was induced by IPTG and identified by SDS- PAGE following purification with affinity chromatography. The results showed that size of Siva1 was 12 kDa, consistent with the molecular weight of the His-Siva1 fusion protein. Functional test demonstrated that Siva1 significantly inhibited the invasion and migration of HCT116 cells. It may thus find clinical application for control of cancers.
Suzhou Industrial Park Center for Disease Control and Prevention, Suzhou, China E-mail : wanzhanzhu@sogou.com.
Ya-Han
YH
Zhang
Lu-Gang
LG
Yu
Wan-Zhan
WZ
Zhu
Sheng-Li
SL
Wang
Dian-Dong
DD
Wang
Yan-Xin
YX
Yang
Xia
X
Yu
D051017:Apoptosis Regulatory Proteins / Q000235:genetics* / Q000302:isolation & purification*; D002465:Cell Movement / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D015179:Colorectal Neoplasms / Q000235:genetics / Q000503:physiopathology; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D005818:Genetic Engineering / Q000379:methods; D045325:HCT116 Cells / Q000166:cytology / Q000502:physiology; D006801:Humans; D010957:Plasmids / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured
D051017:Apoptosis Regulatory Proteins; D011993:Recombinant Fusion Proteins; C106549:SIVA1 protein, human