CAJ | 학술논문

噬菌体展示肽库是一种被广泛用于抗原表位鉴定,结合蛋白筛选的技术。该试验利用噬菌体展示技术筛选与猪呼吸与繁殖障碍综合征病毒（PRRSV）ORF1B复制酶蛋白相互作用的蛋白,并进一步验证筛选蛋白的抗病毒作用。以表达纯化的PRRSV ORF1B蛋白CTD包被高亲和性96孔板作为靶蛋白,应用T7噬菌体展示技术对随机12肽库cDNA文库进行筛选,并分析测序筛选的克隆。将筛选出的克隆序列合成后,在体外验证合成多肽的抗PRRSV效果。结果表明,在4轮的噬菌体筛选后,共得到87个阳性克隆,经测序鉴定出11个筛选的12肽,并通过体外抗病毒试验得到P10是一个具有高抗病毒活性的12肽。试验结果为PRRSV的抗病毒研究奠定了基础。
서균체전시태고시일충피엄범용우항원표위감정,결합단백사선적기술。해시험이용서균체전시기술사선여저호흡여번식장애종합정병독（PRRSV）ORF1B복제매단백상호작용적단백,병진일보험증사선단백적항병독작용。이표체순화적PRRSV ORF1B단백CTD포피고친화성96공판작위파단백,응용T7서균체전시기술대수궤12태고cDNA문고진행사선,병분석측서사선적극륭。장사선출적극륭서렬합성후,재체외험증합성다태적항PRRSV효과。결과표명,재4륜적서균체사선후,공득도87개양성극륭,경측서감정출11개사선적12태,병통과체외항병독시험득도P10시일개구유고항병독활성적12태。시험결과위PRRSV적항병독연구전정료기출。
The phage display peptide library is a widely-used epitope identification technology,which is combined with the protein screening. Phage display technique was used to screen out the proteins interacted with ORF1B copy enzyme protein of porcine respiratory and reproductive syndrome virus （PRRSV） . And the antiviral activity of the screening protein was further verified. The purified CTD protein of PRRSV ORF1B that coated high-affinity 96T board was used as the target protein. T7 phage display technology was applied to screen out the cDNA library of random 12-peptide library. And the screened clones were sequenced and analyzed. After the synthesis of the screened clone sequence, the anti-PRRSV effect of the synthetic polypeptide was verified in vitro. The results showed that 87 positive clones were obtained after 4 circles of phage screening. 11 screened 12-peptide were identified after sequencing. Through in vitro antiviral experiment, it was confirmed that P10 was a 12-peptide with high-antiviral activity. These results laid the foundation for the antiviral research of PRRSV.