發(fā)布時(shí)間：2018-02-23 22:04

  本文關(guān)鍵詞： 副豬嗜血桿菌型 自殺性質(zhì)粒pRE galE基因缺失株　出處：《中國獸醫(yī)科學(xué)》2017年02期 　論文類型：期刊論文

【摘要】：為構(gòu)建副豬嗜血桿菌5型galE基因缺失株,本研究通過重疊PCR構(gòu)建了galE基因的上、下游同源臂,然后將其連接到pRE112載體而構(gòu)建了重組自殺性質(zhì)粒pRE112ΔgalE;使重組質(zhì)粒pRE112ΔgalE轉(zhuǎn)化大腸桿菌X7213,與副豬嗜血桿菌接合轉(zhuǎn)移,應(yīng)用兩步法篩選galE基因缺失株HPS-YA-008ΔgalE,用PCR方法驗(yàn)證galE基因的缺失突變。在此基礎(chǔ)上進(jìn)一步研究galE基因缺失株HPS-YA-008ΔgalE的生長(zhǎng)特性、藥物敏感性、對(duì)保育豬的毒力等生物學(xué)特性。結(jié)果,副豬嗜血桿菌血清5型galE基因缺失株HPS-YA-008ΔgalE構(gòu)建成功,且缺失株的毒力已經(jīng)被致弱,對(duì)藥物的敏感性增加。本試驗(yàn)的研究結(jié)果為研制更加安全的副豬嗜血桿菌弱毒疫苗提供了實(shí)驗(yàn)材料。
[Abstract]:In order to construct the deletion strain of galE gene of Haemophilus parais type 5, the upstream and downstream arms of galE gene were constructed by overlapping PCR. Then the recombinant suicide plasmid pRE112 螖 galE was constructed by ligating it into pRE112 vector, and the recombinant plasmid pRE112 螖 galE was transformed into Escherichia coli X7213. A two-step method was used to screen HPS-YA-008 螖 galE, and PCR method was used to verify the deletion mutation of galE gene. On the basis of this, the growth characteristics, drug sensitivity, virulence of galE gene deletion strain HPS-YA-008 螖 galE, and so on, were further studied. Haemophilus paraswine serotype 5 galE gene deletion strain HPS-YA-008 螖 galE was successfully constructed, and the virulence of the deletion strain was weakened. The results of this study provided experimental materials for the development of a more safe attenuated Haemophilus parasuis vaccine.
【作者單位】： 四川農(nóng)業(yè)大學(xué)動(dòng)物醫(yī)學(xué)院動(dòng)物檢疫實(shí)驗(yàn)室;
【基金】：十二五”國家科技支撐計(jì)劃項(xiàng)目(2013BAD12B04)
【分類號(hào)】：S852.61

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本文編號(hào)：1527782