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Title 

Transgenic tobacco culture cells expressing spike protein gene of porcine epidemic diarrhea virus

 

돼지 유행성 설사병 바이러스 스파크 단백질 유전자 발현 형질전환 담배 배양세포

Authors 

Kyoung-Sil YangH S KimSuk Yoon KwonSang Soo KwakHaeng Soon Lee

Publisher 

Korea Society of Plant Biotechnology

Issue Date 

2008

Citation 

Journal of Plant Biotechnology, vol. 35, no. 1, pp. 87-94

Abstract 

Porcine epidemic diarrhea virus (PEDV) is an infectious and highly contagious virus of swine. In order to develop the transgenic tobacco culture cells producing PEDV antigen protein, four vectors expressing PEDV spike protein (SP) gene under the control of a CaMV 35S promoter were constructed. Four fragments of the SP region of PEDV, SP1 (444 bp, 1487-1930 bp), SP2 (1.7 kb, 2300-3987 bp), SP3 (1.4 kb, 1559-2950 bp), and SP4 (2.6 kb, 9-2643 bp) were amplified by PCR and then C-MYC tag was fused to the end of each SP gene, respectively. These cassettes are inserted into the pCAMBIA2300 (named as 35S::SP1-M, 35S::SP2-M 35S::SP3-M, and 35S::SP4-M, respectively). Tobacco (cv. BY-2) cultured cells were transformed by co-cultivation with Agrobacterium tumefaciens harboring expression vector. We selected kanamycin-resistant calli and checked for the presence of the introduced SP gene using PCR, resulting 70% of them showed the foreign gene. We selected the lines with high-level expression of PEDV antigen protein based on dot blot analysis. Southern blot analysis confirmed that the PEDV SP gene was integrated into the genome of the tobacco cultured cells. Northern blot analysis showed that the introduced gene was highly expressed in transgenic cultured cells. Transgenic tobacco cultured cells-derived antigen induced immunogenicity in mice as determined by a plaque reduction neutralization assay. These results suggest that the vectors expressing PEDV spike protein gene in this study will be useful for the development of transgenic plants and cultured cells producing PEDV antigene protein.

ISSN 

1229-2818

Link 

http://dx.doi.org/10.5010/JPB.2008.35.1.087

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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