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

doi:10.7668/hbnxb.2016.02.014

Abstract

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

CLC Number:

Q78
S432.4+1

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.

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References

[1] Fingerut E,Gutter B,GalliliG,et al.A subunit vaccine against the adenovirus egg-drop syndrome using part of its fiber protein[J].Vaccine,2003,21(21):2761-2766.
[2] Gutter B,Fingerut E,Gallili G,et al.Recombinant egg drop syndrome subunit vaccine offers an alternative to virus propagation in duck eggs[J].Avian Pathology,2008,37(1):33-37.
[3] Tomko R P,Johansson C B,Totrov M,et al.Expression of the adenovirus receptor and its interaction with the fiber knob[J].Experimental Cell Research,2000,255(1):47-55.
[4] 郭建军,范汉东,杨一兵.减蛋综合征病毒纤维蛋白主要抗原结构域原核表达及抗原性鉴定[J].畜牧与兽医,2012,44(2):22-25
[5] 郑金,陈滔,陆吉虎,等.减蛋下降综合征病毒重组KnobS基因在杆状病毒系统中的表达及其免疫原性鉴定[J].江苏农业学报,2015,31(3):600-603.
[6] 史达,宋岩,李一经.乳酸乳球菌作为黏膜免疫活载体疫苗传递抗原的研究进展[J].微生物学报,2006,46(4):680-683.
[7] 李超,王春凤,杨桂连.乳酸菌胞外多糖肠道黏附及免疫调节作用研究进展[J].食品科学,2014,35(11):314-318.
[8] 董懿樱,陈臣,任婧,等.乳酸菌对肠免疫调节功能研究进展[J].中国微生态学杂志,2014,26(2):221-224.
[9] Mohammed B E D,Gahan C G M,Griffin B T.Lactococcus lactis as a cell factory for delivery of therapeutic proteins.[J].Current Gene Therapy,2010,10(1):34-45.
[10] 王相清.共表达猪细小病毒VP2与大肠杆菌不耐热肠毒素B亚单位重组乳酸杆菌构建及其免疫学初步评价[D].哈尔滨:东北农业大学,2009.
[11] Wells J M,Mercenier A.Mucosal delivery of therapeutic and prophylactic molecules using lactic acid bacteria[J].Nat Rev Microbiol,2008,6(5):349-362.
[12] Douglas G L,Klaenhammer T R.Directed chromosomal integration and expression of the reporter gene gusA3 in Lactobacillus acidophilus NCFM[J].Appl Environ Microbiol,2011,77(20):7365-7371.
[13] 郭建军,袁林,曾静,等.干酪乳杆菌Lactobacillus casei 393 upp基因突变株的构建[J].江西科学,2015,33(03):308-311.
[14] 郭建军,龚小华,袁林,等.干酪乳杆菌同源整合载体的构建与鉴定[J].江西科学,2015,33(04):491-494,518.
[15] García-Fruitós E.Lactic acid bacteria:a promising alternative for recombinant protein production[J].Microbial Cell Factories,2012,11(1):1-3.
[16] 宋佰芬.产气荚膜梭菌α毒素基因在干酪乳杆菌基因组中的表达及免疫效果评价[D].哈尔滨:东北农业大学,2014
[17] Goh Y J,Azcarate-Peril M A,O'Flaherty S,et al.Development and application of a upp-based counterselective gene replacement system for the study of the S-layer protein SlpX of Lactobacillus acidophilus NCFM[J].Appl Environ Microbiol,2009,75(10):3093-3105.
[18] Kristich C J,Manias DA,Durmy GM.Development of a method for markerless genetic exchange in Enterococcus faecalis and its use in construction of a srtA mutant[J].Appl Environ Microbiol,2005,71(10):5837-5849.
[19] 杨奇.大肠杆菌DH5α upp基因的敲除及其应用研究[D].南京:南京理工大学,2013.
[20] 宋丽.乳酸乳球菌upp基因缺陷互补型质粒载体系统的构建及其缺失菌生物学特性研究[D].哈尔滨:东北农业大学,2015.

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