重组减蛋综合征病毒<em>KnobS</em>干酪乳杆菌的构建与表达

doi:10.7668/hbnxb.2016.02.014

华北农学报 ›› 2016, Vol. 31 ›› Issue (2): 81-86. doi: 10.7668/hbnxb.2016.02.014

重组减蛋综合征病毒KnobS干酪乳杆菌的构建与表达

郭建军, 袁林, 曾静, 邱小忠, 付锦楠

江西省科学院微生物研究所, 江西南昌 330096

收稿日期:2016-02-15 出版日期:2016-04-28
通讯作者: 袁林(1980-),男,江西德安人,助理研究员,博士,主要从事基因工程与分子生物学研究。
作者简介:郭建军(1984-),男,江西崇仁人,助理研究员,硕士,主要从事有益微生物分子遗传及资源利用研究。
基金资助:
江西省科技支撑计划项目(20142BBF60036);江西省自然(青年)科学基金项目(2013BAB214014)

The Construct and Expression of KnobS Gene of Egg drop syndrome virus in L.casei Genome

GUO Jianjun, YUAN Lin, ZENG Jing, QIU Xiaozhong, FU Jinnan

Institute of Microbiology, Jiangxi Academy of Sciences, Nanchang 330096, China

Received:2016-02-15 Published:2016-04-28

摘要/Abstract

摘要： 以干酪乳杆菌为传递载体,以开发食品级安全的减蛋综合征病毒疫苗为目标,构建了一个温度敏感型自杀型质粒系统pORZP-UKD。将该质粒电转化到干酪乳杆菌L.casei中,经过2次升温处理和5-氟尿嘧啶抗性筛选,挑选阳性克隆,采用PCR和SDS-PAGE方法进行鉴定。KnobS基因整合到干酪乳酸杆菌基因组内,并实现融合蛋白KnobS的分泌表达。表明得到了一株具有无任何选择性标记、携带有KnobS表达盒元件的基因组整合的重组干酪乳酸杆菌KnobSΔupp L.casei,且整合的外源基因能够随着细菌的复制而进行表达,为减蛋综合征病毒免疫保护性抗原KnobS疫苗的进一步研制提供了试验数据。

关键词: 干酪乳杆菌, 减蛋综合征病毒KnobS蛋白, 基因组表达

Abstract: To develop an EDSV(Egg drop syndrome virus) vaccine that live up to the standard of food safety, a temperature-sensitive suicide plasmid pORZP-UKD was constructed using L.casei as a transmitted vector.After transforming it into L.casei using electric shock, two temperature-elevating treatments and a 5-FU screening were performed to get positive clone.Upon the identification of putative clone by PCR and SDS-PAGE, we successfully obtained a recombinant strain, KnobSΔupp L.casei, which has no selective-marker but harbors a KnobS expression box.Any exogenous gene can be integrated into this recombinant strain and express its products as bacteria replicate itself, thus providing the experimental basis for the further development of protective Knobs antigen vaccine against Egg drop syndrome virus.

Key words: Lactobacillus casei, KnobS protein of Egg drop syndrome virus, Genome expression

中图分类号:

Q78
S432.4+1

郭建军, 袁林, 曾静, 邱小忠, 付锦楠. 重组减蛋综合征病毒KnobS干酪乳杆菌的构建与表达[J]. 华北农学报, 2016, 31(2): 81-86. doi: 10.7668/hbnxb.2016.02.014.

GUO Jianjun, YUAN Lin, ZENG Jing, QIU Xiaozhong, FU Jinnan. The Construct and Expression of KnobS Gene of Egg drop syndrome virus in L.casei Genome[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(2): 81-86. doi: 10.7668/hbnxb.2016.02.014.

http://www.hbnxb.net/CN/Y2016/V31/I2/81

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