【摘要】：為了對BVDV-1 P7蛋白進(jìn)行原核表達(dá)及生物信息學(xué)分析,采用RT-PCR的方法擴(kuò)增BVDV NADL株BVDV-1 P7的ORF,擴(kuò)增產(chǎn)物經(jīng)Bam H I、Sal I雙酶切后插入原核表達(dá)載體p EGX4T-1(+),構(gòu)建P7基因原核表達(dá)載體并將其轉(zhuǎn)化至工程菌E.coli BL21(DE3)中進(jìn)行誘導(dǎo)表達(dá),并對其進(jìn)行生物信息學(xué)預(yù)測。結(jié)果表明,成功擴(kuò)增并構(gòu)建了BVDV NADL株的BVDV-1 P7原核表達(dá)載體,命名p EGX4T-1-P7。SDS-PAGE和Western blot表明,在分子質(zhì)量34 k Da的位置出現(xiàn)與預(yù)期大小一致的蛋白條帶;生物信息學(xué)結(jié)果表明在P7蛋白有信號肽,存在疏水區(qū)。該研究成功的表達(dá)了P7蛋白,該蛋白是一個(gè)疏水性蛋白,為其進(jìn)一步的功能研究奠定了堅(jiān)實(shí)的基礎(chǔ)。
[Abstract]:For prokaryotic expression and bioinformatics analysis of BVDV-1 P7 protein, The ORF, amplification product of BVDV-1 P7 of BVDV NADL strain was amplified by RT-PCR and inserted into prokaryotic expression vector p EGX4T-1 (), after being digested with Bam H Il-Sal I and then transformed into E.coli BL21 (DE3) to induce expression. The bioinformatics prediction was carried out. The results showed that the prokaryotic expression vector of BVDV-1 P7 of BVDV NADL strain was successfully amplified and constructed. The identification of p EGX4T-1-P7.SDS-PAGE and Western blot showed that the protein bands were consistent with the expected size at the position of 34 k Da. Bioinformatics results showed that P7 protein had signal peptide and hydrophobic region. This study successfully expressed P7 protein, a hydrophobic protein, which laid a solid foundation for further functional study.
【作者單位】： 黑龍江八一農(nóng)墾大學(xué)動物科技學(xué)院;
【基金】：黑龍江省青年科學(xué)基金(QC2015031) 獸醫(yī)生物技術(shù)國家重點(diǎn)實(shí)驗(yàn)室開放課題基金(SKLVBF201508)
【分類號】：S852.65

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