本文選題：綿羊肺炎支原體　切入點(diǎn)：p基因　出處：《西南農(nóng)業(yè)學(xué)報(bào)》2017年05期 　論文類型：期刊論文

【摘要】：【方法】利用PCR方法對綿羊肺炎支原體(Mycoplasma ovipneumoniae,MO)新疆分離株MO-XJ p74基因進(jìn)行擴(kuò)增、克隆及測序,分析其分子特征。將P74蛋白抗原集中區(qū)編碼序列亞克隆至表達(dá)載體p ET-32a(+),構(gòu)建重組表達(dá)載體p ET-32a(+)-p74C,轉(zhuǎn)化至E.coli BL21(DE3)感受態(tài)細(xì)胞中,IPTG誘導(dǎo)表達(dá)�！窘Y(jié)果】MO-XJ p74基因全長2016 bp,編碼671個(gè)氨基酸;對編碼的氨基酸序列分析發(fā)現(xiàn),該蛋白不含信號(hào)肽序列,9~31位氨基酸序列含有1個(gè)跨膜區(qū),有16個(gè)N-糖基化位點(diǎn)、7個(gè)N-�；稽c(diǎn)、17個(gè)酪蛋白激酶Ⅱ磷酸化位點(diǎn)及5個(gè)蛋白激酶C磷酸化位點(diǎn)。同源性分析顯示MO-XJ P74蛋白氨基酸序列與標(biāo)準(zhǔn)株MO-SC01氨基酸序列同源率為86.5%,其羧基端為P74蛋白抗原集中區(qū)。SDS-PAGE檢測結(jié)果顯示,表達(dá)的P74蛋白抗原表位集中區(qū)片段對分子質(zhì)量約為35.5 k Da,與理論值相符;Western blot分析表明重組P74蛋白具有較強(qiáng)的反應(yīng)原性。【結(jié)論】本研究為進(jìn)一步篩選MO亞單位疫苗及血清學(xué)診斷的候選抗原奠定了前期基礎(chǔ)。
[Abstract]:[methods] the MO-XJ p74 gene of Mycoplasma ovis pneumoniae MOA Xinjiang isolate was amplified, cloned and sequenced by PCR. By analyzing its molecular characteristics, the coding sequence of P74 antigen concentration region was subcloned into the expression vector pET-32a( pET-32a), and the recombinant expression vector pET-32a (pET-32a) was transformed into E. coli BL21DE3. [results] the full-length MO-XJ p74 gene was expressed in 166bp. Encoding 671 amino acids; The amino acid sequence analysis showed that the protein contained a transmembrane region without a signal peptide sequence. There were 16 N-glycosylation sites, 7 N-acylation sites, 17 casein kinase 鈪，

本文編號(hào)：1620026