水貂阿留申病毒NS1基因在大肠杆菌中的分段表达

Segmented Expression of Aleutian Disease Virus (ADV) NS1 Gene Fragment in E.coli

LIU Hong-na¹, WANG Wen-yu¹, ZHANG Yun¹, YANG Yu-hang¹, HAO Jun-wei¹, SHI Kun², LI Jian-ming², DU Rui²

(1.College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China; 2.College of Chinese Medicinal Material, Jilin Agricultural University, Changchun 130118, China)

Received:2013-11-19 Online:2014-04-20 Published:2014-05-27

Abstract

Abstract: We predicted the non-structural protein NS1 of aleutian disease virus (ADV) by DNAStar software Protean module. Then three fragments of NS1 gene named as L1N1, L2N2 and L3N3 were amplified using the laboratory saved NS1-pMD18-T plasmid as template. We constructed three cloning plasmids pMD18-T-L1N1, pGEM-T-L2N2 and pMD18-T-L3N3. After identification, they were linked to pGEX-4T-1 prokaryotic expression vector by two enzymes EcoRⅠ and SalⅠ, respectively, and transformed into E.coli BL21 (DE3) inducing with IPTG. The expressed recombinant proteins were analyzed by SDS-PAGE and Western blotting. The results showed that three recombination prokaryotic expression plasmids pGEX-4T-L1N1, pGEX-4T-L2N2 and pGEX-4T-L3N3 were successfully constructed and the SDS-PAGE results showed that three purpose proteins were expressed about 57, 55 and 44 ku as expected. Furthermore, the proteins were recognized by ADV positive sera and had the reactionogenicity. In the study, the NS1 gene of ADV was expressed segmentedly in E.coli for the first time and laid a foundation for the further study of ADV subunit vaccines.

Key words: aleutian disease virus NS1 gene; E.coli; segmented expression

LIU Hong-na, WANG Wen-yu, ZHANG Yun, YANG Yu-hang, HAO Jun-wei, SHI Kun, LI Jian-ming, DU Rui. Segmented Expression of Aleutian Disease Virus (ADV) NS1 Gene Fragment in E.coli[J]. .

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Segmented Expression of Aleutian Disease Virus (ADV) NS1 Gene Fragment in E.coli

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