本文選題：產(chǎn)腸毒素大腸桿菌 + mSTa-LTB-STb融合蛋白��； 參考：《中國預(yù)防獸醫(yī)學(xué)報(bào)》2017年12期

【摘要】：為獲得產(chǎn)腸毒素大腸桿菌(ETEC)抗原STa突變體(mSTa)、LTb和STb融合蛋白在乳酸菌表達(dá)系統(tǒng)中組成型分泌表達(dá),本研究將mSTa、LTb、STb三價(jià)抗原融合基因,克隆于p23啟動(dòng)子-USP45分泌信號(hào)肽之后,插入到乳酸乳球菌表達(dá)載體pTX8048中,構(gòu)建了重組表達(dá)載體pTX-sls,將其電轉(zhuǎn)化到宿主菌乳酸乳球菌L.lactisNZ9000中進(jìn)行表達(dá),應(yīng)用westernblot、ELISA方法鑒定蛋白表達(dá)情況。將重組乳酸乳球菌pTX-sls/NZ9000口服免疫BALB/c小鼠,分別測(cè)定了免疫后不同時(shí)間血清中特異性IgG、糞便中特異性的sIgA水平以及血清的中和活性;采用MTT法檢測(cè)免疫小鼠脾淋巴細(xì)胞增殖情況,流式細(xì)胞術(shù)檢測(cè)Th細(xì)胞免疫類型。結(jié)果顯示,目的蛋白mSTa-LTB-STb以分泌形式組成型表達(dá),可被陽性血清所識(shí)別。在免疫小鼠血清和糞便中均可檢測(cè)到特異性IgG、sIgA,血清抗體具有一定的中和毒素作用。結(jié)果表明,該重組乳酸乳球菌具有作為口服疫苗的潛在應(yīng)用價(jià)值,本研究為研制ETEC乳酸菌活載體口服疫苗奠定了基礎(chǔ)。
[Abstract]:In order to obtain the constitutive secretory expression of STa mutants of enterotoxigenic Escherichia coli (STa) and STb fusion protein in lactic acid bacteria expression system, the fusion gene was cloned into the signal peptide secreted by p23 promoter -USP45. The recombinant expression vector pTX-slswas constructed by inserting into the expression vector pTX8048 of Lactococcus lactis. The recombinant expression vector pTX-slswas transformed into the host strain of Lactococcus lactis L.lactisNZ9000 for expression. Western blottELISA was used to identify the expression of pTX-sls. BALB/c mice were immunized orally with recombinant Lactococcus lactis pTX-sls/NZ9000. The specific sIgA levels in serum, the specific sIgA level in feces and the neutralization activity of serum were determined at different time after immunization, and the proliferation of spleen lymphocytes in immunized mice was detected by MTT method. Th cell immune type was detected by flow cytometry. The results showed that the target protein mSTa-LTB-STb was expressed in secretory form and could be recognized by positive serum. Specific IgG sIgA can be detected in sera and feces of immunized mice, and serum antibodies can neutralize toxins to some extent. The results showed that the recombinant Lactococcus lactis had potential application value as oral vaccine. This study laid a foundation for the development of oral vaccine of ETEC lactobacillus live vector.
【作者單位】： 東北農(nóng)業(yè)大學(xué)動(dòng)物醫(yī)學(xué)院;
【基金】：國家自然科學(xué)基金(31101845)
【分類號(hào)】：S852.61

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 吳紅,劉晶星;大腸桿菌LT及LTB在免疫預(yù)防小鼠幽門螺桿菌感染中的佐劑性[J];國外醫(yī)學(xué).預(yù)防.診斷.治療用生物制品分冊(cè);2001年01期

2 周素芳,黃秉仁;Ig融合蛋白的應(yīng)用研究進(jìn)展[J];中國生物工程雜志;2002年03期

3 王璋瑜;;蛋白設(shè)計(jì)實(shí)驗(yàn)室公司買下融合蛋白技術(shù)實(shí)用權(quán)[J];生物技術(shù)通報(bào);1990年10期

4 張毅,屈賢銘,楊勝利;融合蛋白基因工程應(yīng)用及研究[J];生物工程進(jìn)展;2000年03期

5 楊林,李國清,黃葆英,王全忠,龍綮新,王s閼，

本文編號(hào)：1926041