鸭疱疹病毒1型的gB蛋白截短表达及其ELISA检测

参照已发表的鸭肠炎病毒（Duck enteritis virus,DEV）基因组序列,利用生物学软件DNAstar进行序列分析,设计并合成一对特异性引物,PCR扩增了长855 bp的gB基因片段.将目的片段克隆至pMD18-T载体,经PCR、酶切及测序鉴定后,定向克隆至pET30a表达载体中,经PCR、酶切鉴定正确后,重组质粒转化BL21表达茵,经IPTG诱导后获得了以包涵体形式表达的重组蛋白.重组蛋白在变性的条件下采用Ni柱亲和层析法纯化,变性蛋白复性后,Western blot检测表明重组蛋白具有良好的反应活性,以该蛋白作为诊断抗原,初步建立了检测鸭肠炎病毒抗体的间接ELISA诊断方法,为鸭肠炎病毒抗体诊断试剂盒的研制奠定了基础.

Truncated Expression, Purification, Activity Detection and Preliminary Application of gB Protein for Duck Herpesvirus Type 1

The fermentation quality and chemical components of the ensiled oat straw and rice straw were investigated in this study. The forage was either ensiled solely or ensiled by mixing the rice straw and oat straw at some rates to determine the optimal mixture between them. The results of this study indicated that the pH value, content of ammonia nitrogen of No. 1 was significantly higher （P〈0.05） than that of other three groups. The pH value and content of ammonia nitrogen of No. 4 was significantly lower than that of other three groups （P〈0.05）. There was significant difference between four groups （P〈0.05）, the crude protein content of No. 4 was the highest. There was no butyric acid in four groups. This study suggested that the directly ensiling oat straw was a better feed, and the effectiveness of the mixed silage which had 70% fresh oat straw was better.