Prokaryotic expression, purification, and polyclonal antibody production of a truncated recombinant rabies virus L protein

Background: rabies virus [RABV] is a deadly neurotropic virus that causes the disease of rabies in humans and animals. L protein is one of the large structural protein of rabies virus, which displays multiple enzymatic activities, and is required for viral transcription and replication

Objectives: a truncated L protein of Rabies virus is being cloned, expressed and purified to produce relevant polyclonal Antibody

Materials and Methods: the gene fragment of L protein of RABV was subcloned into prokaryotic expression vector pET- 28a and transformed into E. coli Rosetta DE3 host strain. The recombinant L protein of RABV was expressed and characterized by SDS-PAGE and western blot analysis using anti-his tag antibody. Mice were immunized with the purified recombinant L protein, the reaction of the anti-serum was checked by immunofluorescence and dot-blot, respectively

Results: the results of PCR and sequencing confirmed that the fragment of L gene of RABV was successfully cloned into the expression vector. The expression of recombinant L protein fragment induced by IPTG was confirmed by the band of 43 kDa in SDS-PAGE and western blot. The antiserum of purified L protein immunized mice was reacted with RABV infected N2a cells and suckling mouse brain tissue lysates

Conclusions: our data showed that the recombinant L protein produced by pET-28a vector was very successful, and the purified L protein could efficiently induce the antibody response in mice. The antiserum could recognize the virus in RABV infected cells and tissue very well

R 692 16 Effect of Diet-Induced Hypercholesterolemia on Kidney of Healthy Rats / 复旦学报（医学版）

Purpose To Investigate the effect of diet-induced hyperchole sterolemia on the kidney ofWistar rats. Methods Male Wistar rats were fed with normal chow supplemented with 5 % cholesteroland observed biochemical changes in plasma lipid concentration, urinary microalbumin excretion, renalfunction, lipid component in renal cortices and morphological changes at 30,60 and 90 days. ResultsTotal plasma cholesterol (TCh) and low density lipoprotein (LDL) concentration were significantly elevatedin the group E (P＜0.05)at 30 days, and progressively increased thereafter, but during the entire study,there ere no differences in plasma urea nitrogen(BUN),creatinine(Cr), and endogenous creatinine clearance(Ccr) between the two groups. Quantitative urinary microalbumin excration was markely elevated in group E( P ＜ 0.05 ). Cholesterol (Ch), phosphatidylcholine(PC) and phosphatidylethanolamine(PE) levels of t he renalcortices were sigificantly increased in the group E at 12 weeks. Progressive development in mesangialhypercellulary, increased mesangial matrix, glomerular capillaries collapes were observed in the group E. Noelectron dense deposits were observed in any of the glomeruli examined. There was a siginificant positivecorrelation for the urinary microalbumin, Ch in the renal corticres, and glomerular size with plasma TCh andLDL concentration. Conclusions The diet-induced hypercholesterolemia may cause lipid nephrotoxicity inWistar rats.