5.Construction of the Expression Vector of Viruslike Particles Containing FMDV IRES RNA
Min DOU ; GuoGuang ZHANG ; GuangFu YU ; HongXin ZHANG ; MingShan SHEN ; Liang CHEN ;
China Biotechnology 2006;0(09):-
The Coat protein and Maturase gene of E.coli bacteriophage MS2 was amplified by PCR,then the gene was cloned into pET32a to construct the intermediate vector pET32aCP.The conservative sequence of FMDV internal ribosome entry site(IRES) was cloned into the downstream of pET32aCP bacteriophage gene to construct the prokaryotic expression vector pCPES.The recombinant plasmid pCPES transformed into E.coli strain BL21(DE3) was induced to express with 1mmol/L IPTG.The expression products were purified by sucrose density gradient centrifugation.The expression products observed by TEM were circular viruslike particles,and the diameter of these particles was about 26nm.The stability of viruslike particles was detected,and the viruslike particles was identified by RTPCR.The results showed that the viruslike particles contain the FMDV IRES RNA and have good stability.The viruslike particles have great prospect as the standard and quality control in the area of RNA virus detection.
9.Effect of daidzein on hepatocellular carcinoma SMMC7721 cell proliferation and tumor stem cell CD133 expression
Min ZHANG ; Hongzhi SUN ; Wei WANG ; Guang BAI ; Xiaodong YU
Chinese Journal of Biochemical Pharmaceutics 2014;(3):28-30
Objective To discuss the effect of daidzein on hepatocellular carcinoma SMMC7721 cell proliferation CD133 expression on tumor stem cell.Methods Hepatocellular carcinoma SMMC7721 cells were cultured,digested and passaged,and divided into six groups with different drug:control group with no daidzein,100 μg/mL daidzein group,200μg/mL daidzein group,300μg/mL daidzein group,400μg/mL daidzein group ,500μg/mL daidzein.The inhibition ratio,hexokinase,alkaline phosphatase and CD133 levels in SMMC7721 cell were detected and compared at 24 h,48 h,72 h among those groups. Results The inhibition ratio was increased by Daidzein dose increasing,and decreased apparently by times extending,especially in 400μg/mL and 500μg/mL daidzeingroups.Compared with control group,the hexokinase and alkaline phosphatase activity and CD133 expression were decreased apparently in groups treated with daidzein(P<0.01).The more the dose,the higher the drop(P<0.01).Conclusion Daidzein can inhibit hepatocellular carcinoma SMMC7721 cell proliferation,and inhibit CD133 expression on tumor stem cell.
10.Expressions of Transferrin Receptor mRNA,Ferritin Receptor mRNA and Iron Regulatory Protein 1 mRNA in the Placentas Complicated with Fetal Iron Deficiency
chun-yan, LIU ; yu-feng, LIU ; shu-guang, ZHANG
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To investigate the expressions of transferrin receptor (TfR) mRNA, ferritin(Fn) mRNA and iron regulatory proteinl (IRP-1) mRNA in the placentas complicated with fetal iron deficiency(ID). Methods Depending on the cord SF,the subjects were divided to fetal ID group and fetal iron sufficient( IS) group. TfR mRNA, Fn mRNA and IRP - 1 mRNA in placentas were measured by RT - PCR. Results 1. The expression of TfR mRNA in ID group was 1.10 ? 0. 26, it was significantly higher than that in IS group (t=0.028 P0.05) ;4. TfR mRNA and Fn mRNA correlated with fetal IS respectively. Conclusion Fetal ID induces highly expressed TfR mRNA and lower level of Fn mRNA to furthest satisfy the iron need for fetus.