本文選題：金黃色葡萄球菌 + 奶牛IgG�。� 參考：《動(dòng)物醫(yī)學(xué)進(jìn)展》2017年10期

【摘要】：為了提高金黃色葡萄球菌(S.aureus)黏附素FnBPB-ClfA基因表達(dá)蛋白對(duì)奶牛的免疫效力,應(yīng)用RT-PCR擴(kuò)增奶牛IgG Fc基因片段與構(gòu)建的金黃色葡萄球菌(S.aureus)FnBPB-ClfA黏附素基因串聯(lián)構(gòu)建質(zhì)粒pMD19T-Fc-FnBPB-ClfA,并通過(guò)雙酶切將其克隆于pET32a(+),以構(gòu)建質(zhì)粒pET32a-Fc-FnBPB-ClfA并進(jìn)行誘導(dǎo)表達(dá)。以鹽酸左旋咪唑(LH)或氫氧化鋁(ALUM)為佐劑,與純化的重組蛋白(bFcFnBPB-ClfA)混合以首次免疫肌肉注射,加強(qiáng)免疫乳頭管灌注或首次免疫、二次免疫及三次免疫均以肌肉注射方式免疫臨近干奶期的奶牛。用間接ELISA和Western blot對(duì)各組牛血清中的抗體進(jìn)行監(jiān)測(cè),評(píng)價(jià)免疫效果。結(jié)果顯示,以LH為佐劑免疫的(A組)牛21d時(shí)抗體水平最高,可達(dá)到1∶1 600;而以ALUM為佐劑免疫的B組和C組抗體水平分別為1∶800和1∶400。IgG水平動(dòng)態(tài)監(jiān)測(cè)結(jié)果表明,以LH為佐劑免疫牛的血清中特異性抗體上升趨勢(shì)明顯高于其他免疫組。研究結(jié)果表明,以首次免疫肌肉注射,加強(qiáng)免疫乳頭管灌注的方式較好,并且bFc-FnBPB-ClfA與LH佐劑混合后免疫奶牛能最有效激發(fā)奶牛免疫反應(yīng)。
[Abstract]:In order to improve the immune effect of the gene expression protein of Staphylococcus aureus (S.aureus) FnBPB-ClfA gene, the IgG Fc gene fragment was amplified by RT-PCR to construct the plasmid pMD19T-Fc-FnBPB-ClfA in series with the constructed Staphylococcus aureus (S.aureus) FnBPB-ClfA adhesion gene, and was cloned to pET32a (+) by double enzyme digestion. The plasmid pET32a-Fc-FnBPB-ClfA was constructed and induced to induce the expression. Using LH or ALUM as the adjuvant, the recombinant protein (bFcFnBPB-ClfA) was mixed with the purified recombinant protein (bFcFnBPB-ClfA) for the first immunization of the intramuscular injection. The immunization of the nipple tube was enhanced or the first immunization was enhanced. The two and three immunizations were immunized near the dry milk by intramuscular injection. The bovine serum antibody in each group was monitored with indirect ELISA and Western blot to evaluate the immune effect. The results showed that the antibody level of bovine 21d with LH as adjuvant was highest and reached 1: 1600, while the level of antibody water in B and C groups with ALUM as adjuvant immunity was 1 to 800 and 1: 400.IgG level dynamic monitoring results. The results showed that the increasing trend of specific antibodies in the serum of cattle immunized with LH was obviously higher than that of other immune groups. The results showed that the method of strengthening the immunization with the immunization of the nipple tube was better by the first immunization of the muscle, and the immunization of cows could be most effectively stimulated by the mixture of bFc-FnBPB-ClfA and LH adjuvant.
【作者單位】： 內(nèi)蒙古農(nóng)業(yè)大學(xué)獸醫(yī)學(xué)院微生物學(xué)與免疫學(xué)實(shí)驗(yàn)室;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(31460664) 農(nóng)牧交錯(cuò)區(qū)生態(tài)型現(xiàn)代奶業(yè)生產(chǎn)模式研究與示范項(xiàng)目(2012BAD12B09-02)
【分類(lèi)號(hào)】：S858.23

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