CAJ | 학술논문

为了更好的研究近两年在我国爆发的猪流行性腹泻（porcine epidemic diarrhea,PED）疫情,本实验室收集不同发病地区的临床样品进行猪流行性腹泻病毒（Porcine epidemic diarrhea,PEDV）RT-PCR检测,并挑选8个阳性样品对其M基因进行扩增,克隆和测序。序列比对的结果显示,8个分离株与14个参考株之间的核苷酸以及氨基酸同源性分别是96.5%～99.9%和96.0%～100%,表明目前流行的PEDV其M基因是相对保守的。同时,我们将ZZ-1株的M基因克隆于原核载体pGEX-6p-1,转入菌株BL21中后获得了大量表达,重组蛋白大小约为50 kDa。利用PEDV阳性猪血清对纯化后的重组M蛋白进行Western blot分析,结果表明重组表达的M蛋白与临床阳性猪血清具有良好的免疫反应性。
위료경호적연구근량년재아국폭발적저류행성복사（porcine epidemic diarrhea,PED）역정,본실험실수집불동발병지구적림상양품진행저류행성복사병독（Porcine epidemic diarrhea,PEDV）RT-PCR검측,병도선8개양성양품대기M기인진행확증,극륭화측서。서렬비대적결과현시,8개분리주여14개삼고주지간적핵감산이급안기산동원성분별시96.5%～99.9%화96.0%～100%,표명목전류행적PEDV기M기인시상대보수적。동시,아문장ZZ-1주적M기인극륭우원핵재체pGEX-6p-1,전입균주BL21중후획득료대량표체,중조단백대소약위50 kDa。이용PEDV양성저혈청대순화후적중조M단백진행Western blot분석,결과표명중조표체적M단백여림상양성저혈청구유량호적면역반응성。
In order to understand recent outbreaks of porcine epidemic diarrhea(PED) in pig populations in China,we collected a few clinical samples from different farms with PED history.A total of 8 isolates were obtained and their M protein gene was sequenced and compared with other 14 reference strains.The nucleotide and amino acid identities among all these strains were 96.5%-99.9% and 96.0%-100%,respectively.The high identity of M gene sequences suggested genetic stability of the PED isolates recently prevalent in pig herds.Then,M gene of ZZ-1 isolate was inserted into pGEX-6p-1 vector that was transformed into BL21 cells.The expressed product was analyzed in SDS-PAGE and Western blotting.The results showed that M protein was expressed with molecular mass of 50 kDa and specifically reacted with PEDV positive swine serum.