問號狀賴型鉤端螺旋體OmpA膜蛋白Loa22基因克隆及表達研究
發(fā)布時間:2018-04-30 03:38
本文選題:鉤端螺旋體 + OmpA基因; 參考:《四川大學》2006年碩士論文
【摘要】:鉤端螺旋體病(簡稱鉤體病)是在世界范圍內(nèi)流行的人獸共患自然疫源性疾病。我國是鉤體病疫情較為嚴重的國家,鉤體病的防治是一項重要的任務。鉤端螺旋體病的預防目前以疫苗為主,但現(xiàn)有疫苗僅能產(chǎn)生短期的、不完全的免疫保護效應,因此對賴型鉤端螺旋體(簡稱鉤體)新型疫苗進行研究,有著重要的現(xiàn)實意義。 細菌的外膜成分由于位置的重要性往往成為疫苗研制的首要選擇。OmpA是一種在腸科桿菌中高度保守的外膜蛋白,它由三個功能性的結構域組成:一個親水的胞外結構域,一個跨膜的β桶狀結構域,一個肽聚糖結構域。OmpA能在無免疫佐劑輔助的情況下誘導特異性的體液和細胞毒性反應,而且還能輔助其它抗原(如Ovalbumin,卵白蛋白)內(nèi)化和交叉遞呈,具有潛在的免疫載體功能,能用于疫苗的研究。我們以賴型鉤體中編碼OmpA膜蛋白的Loa22基因為對象,對其進行克隆分析,并對其進行原核及真核表達。 在本研究中,,我們首先以問號狀賴型鉤體017株、56601株及雙曲鉤體Patoc Ⅰ株基因組為模板PCR擴增目的基因。隨后在原核表達方面,我們構建了Loa22基因與質粒pGEX-4T-1的原核重組表達質粒
[Abstract]:Leptospirosis (Leptospirosis) is a worldwide epidemic of zoonotic diseases. Leptospirosis is a serious epidemic situation in China, and the prevention and control of leptospirosis is an important task. The main prevention of leptospirosis is vaccine, but the existing vaccine can only produce short-term and incomplete immune protection effect. Therefore, it is of great practical significance to study the new vaccine of Leptospira Leptospira (Leptospira Leptospira). The outer membrane components of bacteria are often the primary choice for vaccine development because of the importance of their location. OmpA is a highly conserved outer membrane protein in Enterobacteriaceae, which consists of three functional domains: a hydrophilic extracellular domain. A transmembrane 尾 barrel domain, a peptidoglycan domain. OmpA can induce specific humoral and cytotoxic responses without immune adjuvant, but also assist in the internalization and cross-presentation of other antigens, such as Ovalbumin (ovalbumin). It has potential immune vector function and can be used in vaccine research. The Loa22 gene encoding OmpA membrane protein in Leptospira lanceolaris was cloned and analyzed for prokaryotic and eukaryotic expression. In this study, we first amplified the target gene by PCR from the genomes of 56601 strain of Leptospira interrogans 017 and Patoc I strain of hyperbolic Leptospira. Then, in prokaryotic expression, we constructed the prokaryotic expression plasmid of Loa22 gene and plasmid pGEX-4T-1.
【學位授予單位】:四川大學
【學位級別】:碩士
【學位授予年份】:2006
【分類號】:R377
【引證文獻】
相關期刊論文 前1條
1 張東霞;羅永文;郭霄峰;;副豬嗜血桿菌OmpA抗原表位預測、克隆及表達[J];中國獸醫(yī)學報;2012年01期
本文編號:1822928
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