Construction of Vector for Expression of Hsp65 and Esat-6 Genes Driven by a Maize Endosperm-specific Promoter

  • LI Jun wu ,
  • WANG Shan shan ,
  • SoNG Dong ,
  • LIU Yan ,
  • HUANG Qing hua
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  • Department of Microbiology and Immunology,Medical College of Jinan University,Guangzhou 510632,China

Received date: 2009-02-28

  Online published: 2014-10-14

Abstract

To construct the plant expression plasmid containing Mycobacterium tuberculosis Hsp65 and Esat26 genes, and transform the recombined vector into Agrobacterium tumerfaciens LBA4404. The fusion DNA fragment of Hsp65 and Esat26 were amplied from pEGHLE by polymerase chain reaction(PCR)were cloned into the vector pCRG. The combine fragment of promoter globulin21 and the target gene HLE which get from doubled enzymes digestion of the recombined plasmid pCRGHLE was inserted into the plant expression vector pCAMBIA1300 which contain the gene bar for herbicide resistance. The recombinant plasmid was analyzed by restriction enzyme digestion and the inserted target genes in the pC1300GHLE were verified by nucleotide sequencing. Then transformed pC1300GHLE vector into Agrobacterium tumerfa2 ciens LBA4404 by electroporation. The binary expression plasmid,which could express HSP65 and ESAT26,was correctly constructed. The recombinant vector containing Mycobacterium tuberculosis Hsp65 and Esat26 genes is constructed suc2 cessfully and transformed into LBA4404,and lays a foundation for further study on its immunity effectiveness against MTB.

Cite this article

LI Jun wu , WANG Shan shan , SoNG Dong , LIU Yan , HUANG Qing hua . Construction of Vector for Expression of Hsp65 and Esat-6 Genes Driven by a Maize Endosperm-specific Promoter[J]. Acta Agriculturae Boreali-Sinica, 2009 , 24(3) : 28 -31 . DOI: 10.7668/hbnxb.2009.03.006

References

[1] Mustaesa S.Developmentofitew vaccines and diagnostic reagents against tuberculosis[J].Molecular Immunology,2002,39:113-119.
[2] Lowrie D B,Tasconr E,Bonatovl D,et al.Therapy of tubercu-losis in mice by DNA vaccination[J].Nature,1999,400:269-271.
[3] 师长宏,范雄林,柏银兰,等.结核分枝杆菌Ag85B-ESAT6融合蛋白在小鼠体内诱导的免疫应答及其保护力[J].第四军医大学学报,2004,25(18):1633-1636.
[4] 师长宏,安家泽,唐小凤,等.结核分枝杆菌MPT64-ESAT6融合蛋白在小鼠内诱导的免疫应答及其保护力[J].第四军医大学学报,2006,27(9):769-771.
[5] Paton D J,Valarcher J F,Bergnmnn I,et al.Selection of foot and mouth disease vaccine strains-a review[J].Rev Sci Tech,2005,24.
[6] Kaufmann S H E.Heat shock proteins and the immune re-sponse[J].Immunol Today,1990,11(1):129-136.
[7] Johansen P,Raynaud C,Yang M,et al.Anti-mycobacterial immunity induced by a singh injection of M.leprae HSP65-encoding plasmid DNA inbiodegradable microparticles[J].Inununol Lett,2003,90(2-3):81-85.
[8] Trajkovic V,Natarajan K,Sharma P.Immunomodulatory ac-tion of mycobacterial secretory proteins[J].Microbes Infect 2004,6(5):513-519.
[9] Lowrie D B,Tascon R E,Colston M J,et al.Towards a DNA vaccine against tuberculosis[J].Vaccine,1994,12:1537-1540.
[10] Ravn P,Demissie A,Eguale T,et al.Human T cell response to the ESAT-6 antigen from Mycobacterium tuberculosis[J].J Infect Dis,1999,179:637-645.
[11] Hongxun S,Vladimir M P,Corey M,et al.Regiona,I but not systemicrecroitment,expansion of dendritic cells by a pluron-ic-formulated Flt3-ligand plasmid with vaccine adjuvant ac-tivity[J].Vaccine,2003,21(26):3019-3020.
[12] 李林,佳飞.转基因植物口服疫苗的免疫机制研究进展[J].国外医学免疫学分册,2004,27(6):354-357.
[13] 郝浩永,尉亚辉.转基因番茄表达121服乙肝疫苗[J].食品科学,2007,28(6):201-204.
[14] 陈珍,陈文莉.转基因植物口服疫苗研究的新进展[J].细胞与分子免疫学杂志,2006,22(6):831-833.
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