禽呼肠孤病毒分离株P10、P17、δ3蛋白基因的表达及抗血清的制备

Expression of P10,P17,δ3 Proteins of Avian Reovirus Field Strain and Preparation of Their Antisera

禽呼肠孤病毒的感染在我国鸡群中非常普遍，它除了可引起鸡传染性关节炎外，还可能引起矮小综合症和免疫抑制等。从鸡群中分离到的不同毒株ARV的毒力差别较大，抗原性也不尽相同。ARV是一种双股RNA病毒，病毒颗粒中的基因组有10个不同的独立节段组成。

Three genes, P10, P17 and δ3, in S1 segment of avian reovirus （ARV） were used as template and amplified by RT-PCR from a field strain DF0401 with primers synthysized according to the published sequences. PCR product of each gene was separately cloned into pGEX-6p-1 down to GST gene for expression of fusion protein. The E. coli BL21 was transformed with positive recombinant expression plasmid. Each selected BL21 was cultured at 37℃ , induced with IPTG to express the fusion protein. Each fusion protein was recovered from gel electrophoresis and then was used to immunize mice. By indirect immunofluorescence assay, the mouse antisera to P10,P17,δ3 gave positive fluorescence in ARV- infected chicken embryo fibroblast cells, the antibody titers reached 26,26, 28 respectively. The results indicated that three proteins P10 P17 δ3 expressed in E. coli still maintained their antigenicity partially, and the antibody titers to three proteins were high enough to be used as monovalent antisera for diagnostic purpose.