Construction Of Eukaryotic Expression Vector Of ALR And Experiment Research On Acute Hepatic Injury In Rats

Objective: Augmenter of liver regeneration (ALR) cloned from liver of weaning rats is a novel hepatic stimulator. It was reported that ALR mRNA expression was correlated with DNA synthesis of hepatocytes after partial hepatectomy, and could significantly promote the survival rate of intoxicated hepatitis rats. As the specific mitogen, ALR could specially enhance hepatocyte proliferation and might play an important role in recovery of liver injury. In this study, a eukaryotic expression vector for ALR fused with enhanced green fluorescent protein (EGFP) was constructed. In the way of gene therapy, pEGFP-C2-ALR was injected into the rats of CCl₄-intoxicated hepatic injury, and then the expression of GFP in the liver tissue and other tissues, hepatic pathology and the recovery of hepatic injury were investigated in rats.Methods: PCR was performed using plasmids pBV220-ALR as templat under the following conditions: 94°C for 5min, then 94°C for 30sec, 50°C for 20sec and 72°C for 10sec for 30 cycles, last 72°C for 10min. The purified ALR fragment digested by EcoR I and Sal I was ligated with efficient eukaryotic expression vector pEGFP-C2 and...