本文選題：密碼子優(yōu)化 + 乙型肝炎　； 參考：《南京醫(yī)科大學學報(自然科學版)》2009年03期

【摘要】：目的:觀察密碼子優(yōu)化能否增強乙型肝炎病毒表面抗原中蛋白核酸疫苗的免疫原性。方法:根據(jù)乙型肝炎病毒(adr亞型)表面抗原中蛋白(MHBs)的氨基酸序列,在不改變其氨基酸序列的基礎上,設計并人工合成了密碼子優(yōu)化的基因,并將該基因克隆到核酸疫苗載體pSW3891中,構建了密碼子優(yōu)化的表達MHBs核酸疫苗(命名為:pSW3891/MHBs/adr/opt,簡稱opt)。用上述核酸疫苗與表達野生型MHBs核酸疫苗(命名為:pSW3891/MHBs/adr,簡稱adr)及空載體質(zhì)粒pSW3891一起瞬時轉(zhuǎn)染293T細胞,應用蛋白質(zhì)印跡法檢測轉(zhuǎn)染細胞中MHBs的表達。采用肌肉注射法,以opt、adr及pSW3891,分別對BALB/c小鼠進行免疫。用ELISA方法檢測免疫后小鼠血清中HBs抗體的滴度,ELISPOT法檢測免疫小鼠表面抗原特異性分泌IFN-γ的脾細胞。結果:opt和adr均可在體外293T細胞中高效表達,且opt的蛋白表達量要高于野生型。opt免疫組小鼠特異性抗體滴度和免疫小鼠表面抗原特異性分泌IFN-γ脾細胞數(shù)量都要顯著的高于adr免疫組小鼠。結論:密碼子優(yōu)化可以增強乙型肝炎病毒DNA疫苗在體外的蛋白表達量及免疫小鼠的體液免疫和細胞免疫原性。
[Abstract]:Aim: to observe whether codon optimization can enhance the immunogenicity of hepatitis B virus surface antigen protein nucleic acid vaccine. Methods: according to the amino acid sequence of the surface antigen of hepatitis B virus (HBV / ADR), a codon optimized gene was designed and synthesized without changing its amino acid sequence. The gene was cloned into nucleic acid vaccine vector pSW3891 and codon optimized MHBs nucleic acid vaccine was constructed. The above nucleic acid vaccine was used to transfect 293T cells with the expressed wild type MHBs nucleic acid vaccine (named as w / pSW3891 / MHBs / adrr) and no-loaded pSW3891. The expression of MHBs in transfected cells was detected by Western blot. BALB/c mice were immunized with optadr and pSW3891 by intramuscular injection. The titer of HBs antibody in serum of immunized mice was detected by ELISA method. The spleen cells secreting IFN- 緯 on the surface antigen of immunized mice were detected by Elispot method. Results both of them were highly expressed in 293T cells. The protein expression of opt was higher than that of wild-type. Opt immunized mice and the number of IFN- 緯 splenocytes secreted by surface antigen of immunized mice was significantly higher than that of adr immunized mice. Conclusion: codon optimization can enhance the protein expression of hepatitis B virus DNA vaccine in vitro and the humoral and cellular immunogenicity of immunized mice.
【作者單位】： 南京醫(yī)科大學第一附屬醫(yī)院感染科;
【基金】：國家自然科學基金資助(30371276)
【分類號】：R392

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