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Construction of CTB-PROIN and CTB-INSB Fusion Gene and Their Expression in Escherichia Coli and Transgenic Plant

Author: ChenLi
Tutor: ZhangDaBing
School: Ningxia University
Course: Biochemistry and Molecular Biology
Keywords: Cholera toxin B subunit Proinsulin Insulin B chain E. coli Transgenic potato
CLC: R346
Type: Master's thesis
Year: 2005
Downloads: 81
Quote: 0
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Abstract


Oral pancreatic islet -specific antigen can induce immune tolerance to prevent the occurrence of spontaneous autoimmune diabetes . More effectively to the insulin protein absorption by the gut-associated lymphoid tissue , the research to build a strong immune adjuvant cholera toxin B subunit (cholera toxin B subunit, CTB) and proinsulin ( proinsulin , PROIN ) and insulin B chain (insulin chian , B the fusion gene INSB ) CTB- PROIN , the CTB - INSB explore the possibility of type 1 diabetes oral vaccine , using potato production research content and results are as follows : First fusion gene CTB- PROIN CTB - INSB was cloned into the E. coli expression vector for PET - 30a ( ) , and induction of its expression . Expressed in E. coli fusion protein the CTB-PROIN and the CTB-INSB with GM1 glycoside lipid binding , and have dual antigen CTB and insulin resistance . On this basis , were constructed containing the potato tuber-specific promoter, patatin , fusion gene, and a potato proteinase inhibitor gene II ( PIN2) of the 3 'end untranslated region (UTR) sequence and the NOS terminator of the two plants transformation vector , transformed by Agrobacterium tumefaciens in potato transgenic potato plants containing the fusion gene . The transgenic plant expression of the fusion protein with cholera toxin and insulin double antigen , ganglioside GM1 binding , with potential biological activity . To take advantage of this immune NOD mice transgenic potato , laid the foundation for further analysis of its immunological properties .

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