馬立克氏病毒meq基因缺失株SC9-1通過自然重組獲得meq能力的分析
發(fā)布時間:2018-01-19 10:41
本文關(guān)鍵詞: 馬立克氏病病毒 疫苗毒株SC- 超強毒株Md meq基因 PCR鑒定 出處:《病毒學報》2017年01期 論文類型:期刊論文
【摘要】:為了探究meq基因缺失的馬立克氏病毒疫苗株SC9-1與超強毒株Md5是否能夠通過自然重組獲得meq基因的能力,將SC9-1疫苗毒和Md5超強毒共同感染雞胚成纖維細胞(CEF),并在CEF上連續(xù)傳三代,提取單個蝕斑的病毒DNA。同時將Md5超強毒接種免疫過SC9-1疫苗株的SPF雞,在不同的時間點分離病毒,提取單個蝕斑的病毒DNA。將兩種方式獲得的病毒DNA進行PCR驗證,并將香啤酒重組酶位點(FRT)殘留序列克隆測序,比較其同源性。兩種方式鑒定的病毒均為SC9-1或是Md5,沒有檢測到重組病毒,而且FRT殘留序列同源性為100%。結(jié)果 SC9-1沒有從野生毒株Md5獲得缺失的meq基因,而且meq基因敲除區(qū)具有很好的遺傳穩(wěn)定性。
[Abstract]:In order to investigate the ability of SC9-1 and Md5, the Marek's virus vaccine strain with missing meq gene, to obtain meq gene by natural recombination. The chicken embryo fibroblasts were co-infected with SC9-1 vaccine virus and Md5 supervirulent virus and passed on CEF for three consecutive generations. A single plaque virus DNA was extracted. At the same time, the SPF chicken vaccinated with SC9-1 vaccine was inoculated with Md5 supervirulent virus, and the virus was isolated at different time points. The single plaque virus DNA was extracted. The virus DNA obtained from two ways was verified by PCR, and the residual sequence of the recombinant enzyme site was cloned and sequenced. Comparing their homology, the two methods were identified as SC9-1 or MD5, and no recombinant virus was detected. The homology of FRT residue sequence was 1000.Results SC9-1 did not obtain the missing meq gene from wild strain Md5, and the meq gene knockout region had good genetic stability.
【作者單位】: 山東農(nóng)業(yè)大學動物醫(yī)學院;
【基金】:國家自然科學基金青年基金項目(項目號:31402235);題目:Ⅰ型馬立克氏病毒獨特基因sorf2生物學功能的研究
【分類號】:S852.65
【正文快照】: 馬立克氏病(Marek’s disease,MD)是由馬立克氏病毒(Marek’s disease virus,MDV)引起的雞的一種腫瘤性疾病[1]。MDV屬于α皰疹病毒,主要分為致病性的Ⅰ型和無致病性的Ⅱ、Ⅲ型MDV[2]。MD是目前唯一可用疫苗預防的禽病毒性腫瘤疾病,近些年來隨著MDV毒力的不斷增強,相繼出現(xiàn)了
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