ABSTRACT
Objective To construct a shuttle plasmid for inducible gene expression in Borrelia burgdorferi (B.burgdorferi) with an advantage of flexible genetic manipulation.Methods The IPTG-inducible lac repressor/operator system from Escherichia coli (E.coli) was adopted and modified in the current study.The plasmid shuttle vector was developed by inserting multiple cloning sites,FLAG and HA tags into the shuttle vector by molecular cloning approaches.The target gene was inserted at the site under the control of the promoter (Tn5 derivate) in plasmid pQE30.This promoter contained two lac operators and a codonoptimized lacI gene driven by flaB promoter.Results A plasmid shuttle vector,pJJ275,was successfully constructed with the ability to express target genes in B.burgdorferi in the presence of IPTG.By using this system,a HA-tagged rpoS gene was introduced into the typical infectious strain B.burgdorferi B31.The target gene expression induced by IPTG was confirmed at transcriptional and translational levels.The RpoS dependent virulence factor of Borrelia,OspC,was also detected,indicating that the expressed protein was functional.Conclusion The constructed plasmid shuttle vector can express exogenous genes in B.burgdorferi with an inducible feature and an advantage of flexible genetic manipulation.It can be applied for genetic manipulation of B.burgdorferi involved in gene regulation and complementation.
ABSTRACT
Objective To evaluate the correlation between high-sensitivity C-reactive protein(hsCRP)and neutrophil alkaline phosphataee(NAP).Methods 74 patients with different hsCRP levels and 20 healthy subjects were selected.hsCRP and NAP were detected in every person,then the t test and correlation analysis were conducted.Results The NAP integral value of high-value hsCRP group and mid-value were significantly higher than the low-value hsCRP group(P ＜ 0.01),and the NAP group average positive rate of the high,median hsCRP values were 81.8％and 82.3％,which were higher than low-value of hsCRP group(53.6％).Compared the correlation between each group of hsCRP value and integral value of NAP,the hsCRP of high-value hsCRP group has a good correlation with NAP integral value(P ＜ 0.01),while the hsCRP of mid-value of hsCRP group and NAP integral is less relevant(P ＞0.05).Conclusion hsCRP as an acute reactive protein exists a good correlation with NAP,when hsCRP ＞ 50mg/L,it can partially replace NAP.